Compositions and methods of making polymerizing nucleic acids

ABSTRACT

Provided herein are compositions and methods of making high density nucleic acid polymers.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a 35 U.S.C. § 371 National Stage application ofInternational Application No. PCT/US15/42482, filed Jul. 28, 2015; whichclaims the benefit of priority under 35 U.S.C. § 119(e) of U.S.Provisional Patent Application No. 62/029,833, filed Jul. 28, 2014 Thedisclosure of each of the prior applications is considered part of, andis incorporated by reference, in the disclosure of this application.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

This invention was made with government support under grant numberW911NF-14-1-0169 awarded by the Department of Defense. The Governmenthas certain rights in the invention.

REFERENCE TO A “SEQUENCE LISTING,” A TABLE, OR A COMPUTER PROGRAMLISTING APPENDIX SUBMITTED AS AN ASCII FILE

The Sequence Listing written in file 48537-559001WO_ST25.TXT, created onJul. 28, 2015, 531 bytes, machine format IBM-PC, MS-Windows operatingsystem, is hereby incorporated by reference.

BACKGROUND

Structures containing nucleic acids decorating surfaces, particles andpolymers can be made. There is evidence that the density of nucleicacids in such arrays imparts special, highly unusual and uniqueproperties on the nucleic acids. Unfortunately, in the case of polymericchemistry, high densities have not been achieved because all techniquesrely on post-polymerization modification (conjugation) of relativelyhigh molecular weight polymers to relatively high molecular weightnucleic acids. There is a need for methods for generating a nucleic acidpolymer wherein every single position of the polymer backbone is anucleic acid as a monomer. The present invention provides this need andprovides related advantages as well.

BRIEF SUMMARY

The present invention is based, in part, on the discovery of a novelcomposition of a polymer, e.g., brush polymer, wherein every position ofthe polymer backbone contains a nucleic acid as a monomer. Thecomposition serves as an informationally encoded synthetic system withan ultra high density of nucleic acids. Also provided herein are methodsfor synthesizing the poly(oligonucleotide).

This is the first example of a nucleic acid-based polymer material witha high density of nucleic acids such that each position contains anucleic acid. Such a material can be used as a nucleic acid deliveryvehicle, a sensor, a probe, or functional nucleic acid-based materialfor other applications, such as for medicine and advanced materials.Potential applications include the facile preparation of materials foraffinity purification of DNA, gene and nucleic acid delivery to cells,and in the development of materials capable of programmingself-assembly. The nucleic acid-based polymer material possesses novelproperties including nanomaterial formation and DNA-binding.

The poly(oligonucleotide) material described herein is entirelydifferent from post-polymerization modified polymeric materials and/ornanoparticle-DNA conjugates and/or DNA-surface conjugates of any otherkind (e.g. graft-onto polymerization). Other examples of nucleicacid-based material involves first preparing a template, and thenmodifying that template. This, due to thermodynamics and kinetics,results in a less than optimized density of nucleic acid, and it is thedensity of the array that gives unique properties including high bindingaffinities and nuclease resistance (a general resistance to degradationthat plagues almost every type of formulation of nucleic acid).Protecting from degradation and packaging nucleic acids such that theyretain biological function is a key requirement in medicines related tonucleic acid based technologies. The discovery of nucleic acid-basedpolymers by the inventors has solved these and other problems.

In one aspect, provided herein is a synthetic nucleic acid polymercomposition including a plurality of nucleic acid monomers and a polymerbackbone, wherein each position of the backbone contains a nucleic acidmonomer. Each nucleic acid monomer can include a natural or an unnaturalnucleic acid. In some instances, the nucleic acid is RNA, DNA, PNA, LNAor any combination thereof. Each nucleic acid monomer can include atleast 5 nucleic acids or more. Each nucleic acid monomer can include atleast 10 nucleic acids or more. The nucleic acid composition can have adegree of polymerization of at least 5 or more. The compositions can bein the form of a brush polymer. In some embodiments, the composition canbe in the form of a micelle. Alternatively, the compositions can be inthe form of a nanoparticle.

In a second aspect, provided herein is a method for or generating asynthetic nucleic acid polymer composition comprising a plurality ofnucleic acid monomers and a polymer backbone, wherein each position ofthe backbone contains a nucleic acid monomer, the method comprisinggraft-through polymerizing a plurality of nucleic acid monomers. Thestep of polymerizing can include using a ROMP initiator. In someembodiments, the composition has a degree of polymerization of at least5 or more.

In an aspect is provided a graft polymer including a linear backbonecovalently bound to a plurality of oligonucleotide branches. The graftpolymer is assembled by graft-through polymerization of a plurality ofoligonucleotide monomers including a polymerizable monomer covalentlybound to an oligonucleotide. The oligonucleotide thereby forms each ofthe plurality of oligonucleotide branches.

In an aspect is provided a block graft copolymer including a linearbackbone covalently bound to a plurality of oligonucleotide branches anda plurality of non-oligonucleotide side chains, wherein: the pluralityof oligonucleotide branches form a first block portion of the graftcopolymer and the non-oligonucleotide side chains form a second blockportion of the graft copolymer; the graft copolymer is assembled bygraft-through polymerization of a plurality of oligonucleotide monomersand a plurality of non-oligonucleotide monomers, wherein each of theplurality of oligonucleotide monomers includes a polymerizable monomercovalently bound to an oligonucleotide, the oligonucleotide therebyforming each of the plurality of oligonucleotide branches; and each ofthe plurality of non-oligonucleotide monomers includes the polymerizablemonomer covalently bound to a non-oligonucleotide moiety, thenon-oligonucleotide moiety thereby forming each of the plurality ofnon-oligonucleotide side chains.

In an aspect is provided an amphiphilic block graft copolymer includinga linear backbone covalently bound to a plurality of oligonucleotidebranches and a plurality of hydrophobic side chains, wherein: theplurality of oligonucleotide branches form a hydrophilic block portionof the amphiphilic graft copolymer and the hydrophobic side chains forma hydrophobic block portion of the amphiphilic graft copolymer; thegraft copolymer is assembled by graft-through polymerization of aplurality of oligonucleotide monomers and a plurality of hydrophobicmonomers, wherein each of the plurality of oligonucleotide monomersincludes a polymerizable monomer covalently bound to an oligonucleotide,the oligonucleotide thereby forming each of the plurality ofoligonucleotide branches; and each of the plurality of hydrophobicmonomers includes the polymerizable monomer covalently bound to ahydrophobic moiety, the hydrophobic moiety thereby forming each of theplurality of hydrophobic side chains.

In an aspect is provided a micelle including an amphiphilic block graftcopolymer described herein, including in an aspect, embodiment, example,figures, table, scheme, or claim as provided herein.

In an aspect is provided a nanoparticle including an amphiphilic blockgraft copolymer described herein, including in an aspect, embodiment,example, figure, table, scheme, or claim as provided herein.

In an aspect is provided a method of making a graft polymer, the methodincluding: (i) reacting a plurality of oligonucleotide monomers with apolymerization catalyst or initiator, wherein each of the plurality ofoligonucleotide monomers includes a polymerizable monomer covalentlybound to an oligonucleotide; and (ii) terminating the reacting with achain terminator or transfer agent.

In an aspect is provided a method of making an amphiphilic block graftcopolymer, the method including: (i) reacting a plurality ofoligonucleotide monomers with a polymerization catalyst thereby forminga hydrophilic block portion, wherein each of the plurality ofoligonucleotide monomers includes a polymerizable monomer covalentlybound to an oligonucleotide; (ii) reacting the hydrophilic block portionwith a plurality of hydrophobic monomers and the polymerization catalystthereby forming the amphiphilic block graft copolymer, wherein each ofthe plurality of hydrophobic monomers includes the polymerizable monomercovalently bound to a hydrophobic moiety.

In an aspect is provided a method of making an amphiphilic block graftcopolymer, the method including: (i) reacting a plurality of hydrophobicmonomers with a polymerization catalyst thereby forming a hydrophobicblock portion, wherein each of the plurality of hydrophobic monomersincludes a polymerizable monomer covalently bound to a hydrophobicmoiety; (ii) reacting the hydrophobic block portion with a plurality ofoligonucleotide monomers and the polymerization catalyst thereby formingthe amphiphilic block graft copolymer, wherein each of the plurality ofoligonucleotide monomers includes the polymerizable monomer covalentlybound to an oligonucleotide.

Other objects, features, and advantages of the present invention will beapparent to one of skill in the art from the following detaileddescription and figures.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1C show synthesis and characterization of apoly(oligonucleotide). FIG. 1A: PNA-norbornyl monomer (PNA-Nb)polymerized using ROMP initiator (IMesH₂)(C₅H₅N)₂(Cl)₂Ru═CHPh (“Ru”) toform poly-PNA homopolymer, I, and poly-PNA block copolymer, II. FIG. 1B:Representative percent conversion for I determined by the disappearanceof the olefin signal associated with PNA-Nb in 1HNMR. FIG. 1C:Representative SEC-MALS for II. M_(n)=28,270 indicating a degree ofpolymerization of 5 for the PNA block.

FIG. 2 shows structures of monomers used for block copolymerpreparation.

FIGS. 3A-3E show that poly-PNA amphiphile II was dialyzed from DMSO intoH₂O to generate nanoparticles. FIG. 3A: DLS data indicating ahydrodynamic radius of 25 nm. FIG. 3B: T_(m) of PNA-NP with acomplementary DNA sequence was found to be 58.1° C. FIG. 3C:Negative-stained TEM of PNA-NP provided evidence of spherical 20 nmdiameter nanoparticles. Sequence legend: GCTCAGTAAA (SEQ ID NO: 1).Atomistic models of (FIG. 3D) II and (FIG. 3E) PNA-NP. II is shown in aconformation present within PNA-NP.

FIGS. 4A-4B provide Scheme 1 which shows known methods for theincorporation of multiple nucleic acids of nucleobases into polymers.FIG. 4A shows Post-polymerization modification of a polymer with anucleic acid.⁵¹⁻⁵³ FIG. 4B: Polymerization of a pyrimidine base as amodified monomer.⁴⁴⁻⁵⁰

FIG. 5. ¹H NMR spectra for I.

FIG. 6. ¹H NMR spectra for III.

FIG. 7. SEC-MALS for I.

FIG. 8. Synthesis of PNA block copolymers using Grubb's 2nd generationmodified catalyst in ROMP.

FIG. 9. ¹H NMR timescale for ROMP of II.

FIG. 10. ¹H NMR timescale for ROMP of 3 identical block copolymer ratiosIV-VI.

FIGS. 11A-D. ¹H NMR timescale for ROMP of VII-X.

FIGS. 12A-12B. (12A) SEC-MALS of II and (12B) Chemical shift.

FIGS. 13A-13D. SEC-MALS of IV-VI (FIGS. 13A-13D).

FIGS. 14A-14B. SEC-MALS of (FIG. 14A) homophenyl block as well as (FIG.14B) SEC-MALS of VII.

FIGS. 15A-15B. SEC-MALS of (FIG. 15A) homophenyl block as well as (FIG.15B) SEC-MALS of VIII.

FIGS. 16A-16B. SEC-MALS of (FIG. 16A) homopeg block as well as (FIG.16B) SEC-MALS of IX.

FIGS. 17A-17B. SEC-MALS of (FIG. 17A) homo quaternary amine block aswell as (FIG. 17B) SEC-MALS of X.

FIGS. 18A-18D. DLS and TEM of PNA-NP made from II.

FIGS. 19A-19C. Tm data for PNA-NP and complementary DNA sequence.

FIGS. 20A-20C. Raw Tm data for PNA-NP and complementary DNA, as well asnon-complementary DNA sequence.

FIGS. 21A-21B. Raw Tm data for PNA-NP without complementary DNA andcomplementary DNA without PNA in PBS.

FIG. 22: Partial Charges on Adenine Base in PNA.

FIG. 23: Partial Charges of Guanine Base in PNA.

FIG. 24: Partial Charges for Thymine Base in PNA.

FIG. 25: Partial Charges of Cytosine Base in PNA.

FIG. 26: Partial Charges for a Unit of the Peptide Chain in PNA.

FIG. 27: Partial Charges for a Unit of the Hydrophobic Chain.

FIG. 28: HPLC Purification of PNA Sequences.

DETAILED DESCRIPTION OF THE INVENTION

In embodiments, the inventors have, inter alia, avoided shortcomingsassociated with post-polymerization modification reactions (e.g.graft-onto polymerization) and have developed nucleic acid brushpolymers and amphiphilic brush copolymers by direct polymerization viagraft-through polymerization of a nucleic acid. Provided herein is thefirst example of a polymer-nucleic acid bioconjugate generated viadirect polymerization of an oligonucleotide monomer. In addition, thesematerials show cooperative hybridization to complementary DNAoligonucleotides.

In embodiments, the methods described herein provide an efficientsynthetic strategy for the incorporation of nucleic acids into particleand polymer-based materials, with potential applications including thefacile preparation of materials for affinity purification of DNA, geneand nucleic acid delivery to cells, and in the development of materialscapable of programmed self-assembly. In embodiments, the covalentincorporation (direct polymerization or graft-through) of peptidenucleic acids into polymer-based nanomaterials could facilitate cellularinternalization of nucleic acid sequences that could then be capable ofregulating mRNA levels in live human cells, while maintaining theenzymatic resistance of non-naturally occurring PNAs. In embodiments,this could occur through gene interference and/or genetically guided orenhanced theranostics.

I. Definitions

The term “nucleic acid polymer” or “poly(oligonucleotide)” refers to atrue polymer of oligonucleotides that contains a polymer backbone with anucleic acid as a monomer at every position of the backbone (i.e., anoligonucleotide (e.g., PNA) connected at one end of the oligonucleotideto the polymer monomer moiety). In embodiments, a“poly(oligonucleotide)” is a polymer of norbornylmonomer-oligonucleotides (e.g., 2-1000-mers) forming a brush polymerwhere each branch is an oligonucleotide (e.g., 2-1000-mer). The termincludes any polymerized nucleic acid containing material generatedthrough direct polymerization of monomers including a nucleic acid.

The term “nucleic acid” includes at least two nucleotides (e.g., naturaland/or synthetic). The nucleotide can include RNA, DNA, PNA, LNA, andother modified natural or unnatural formulations of a nucleotide. Theterm “nucleotide” as used herein refers to a single nucleotide, and theterms “nucleic acid,” “polynucleotide,” and “oligonucleotide” refer tothe plural thereof.

The term “nucleobase” is used in accordance with its meaning inmolecular biology and genetics and refers to a nitrogen-containingbiological compound (nitrogenous base) that may be linked to a polymerbackbone (e.g. an amide backbone as in peptide nucleic acids or a sugarwithin nucleosides as in DNA and RNA. The nitrogenous bases arewell-known in the art and include, for example the naturally occurringcytosine, guanine, adenine, thymine, uracil as well as natural andnon-natural derivatives thereof. In embodiments, to the nitrogenous baseis cytosine, guanine, adenine, thymine, or uracil. In embodiments, thenitrogenous base is hypoxanthine, xanthine, 7-methylguanine,5,6-dihydrouracil, 5-methylcytosine, and/or 5-hydroxymethylcytosine. Inembodiments, the nucleobase is attached to a detectable moiety. Inembodiments, the nucleobase includes an isomer of a naturally occurringnitrogenous base such as isoguanine or isocytosine. In embodiments,nucleobases are linked through a nucleic acid backbone such as a DNA orRNA backbone or peptide nucleic acid backbone. Internucleotide linkagesmay include one or more of phosphodiester, phosphoramidate,phosphorodiamidate, phosphorothioate (also known as phosphothioate),phosphorodithioate, phosphonocarboxylic acids, phosphonocarboxylates,phosphonoacetic acid, phosphonoformic acid, methyl phosphonate, boronphosphonate, amide and/or O-methylphosphoroamidite linkages.

The term “nucleic acid monomer” refers to a monomeric unit (e.g., singleunpolymerized precursor to a polymer or single repeated unit within apolymer (where each repeated unit is identical within the length of thepolymer but may vary in sidechains (e.g., branches) connected to therepeated unity) containing more than one nucleotide joined by one ormore bonds.

The term “degree of polymerization” refers to the number of monomericunits in a polymer molecule.

The abbreviations used herein have their conventional meaning within thechemical and biological arts. The chemical structures and formulae setforth herein are constructed according to the standard rules of chemicalvalency known in the chemical arts.

The term “graft-through polymerization” is used in accordance with itsplain meaning in the art of polymer chemistry and refers to amacromonomer method of graft polymer (e.g., polymer or copolymer)synthesis. The method forms the graft polymer (also referred to in thepolymer chemistry arts as the macromolecule) by polymerizingmacromonomer molecules having one polymerizable monomer end-group whichenables it to act as a monomer molecule, contributing a single monomericunit to a chain of the final macromolecule (graft polymer). Unlike graftpolymers assembled by graft-to (also commonly referred to as“graft-onto”) polymerization, graft polymers assembled by graft-throughpolymerization do not include unreacted functional groups (or chemicalvestiges thereof) within the linear backbone used to graft side chainpolymers to the linear backbone. Graft-through polymerization includesmethods of polymerization for synthesizing a polymer with monomer sidechains (e.g., nucleic acids) using a known polymerization strategyamenable to the functional groups involved, including protected andunprotected forms. Graft-through polymerization may include synthesizinga polymer with monomer side chains (e.g., nucleic acid monomers) whereinthe monomer side chains are not further derivatized or modified aftersynthesis of the polymer. Thus, graft polymers assembled bygraft-through polymerization include high density polymers havingsmaller distances between side chains relative to graft polymersassembled by graft-to polymerization. In embodiments, the graft polymeris a brush polymer. In embodiments, the graft polymer synthesis is abrush polymer synthesis. In embodiments, the graft-throughpolymerization employs atom transfer radical polymerization (ATRP),ring-opening metathesis polymerization (ROMP), anionic and cationicpolymerizations, free radical living polymerization, radiation-inducedpolymerization, ring-opening olefin metathesis polymerization,polycondensation reactions, or iniferter-induced polymerization.

The term “brush polymer” is used in accordance with its meaning in theart of polymer chemistry and refers to a layer of polymers (e.g.,sidechains, oligonucleotides) attached with one end to a common support(e.g., polymer, linear polymer, surface) The brush polymer may becharacterized by the high density of grafted chains (e.g.oligonucleotides, sidechains). The limited space may lead to a strongextension of the chains and unique properties of the system.

The term “polymerizable monomer” is used in accordance with its meaningin the art of polymer chemistry and refers to a compound that maycovalently bind chemically to other monomer molecules to form a polymer.An example of a polymerizable monomer is a ROMP polymerizable monomer,which is a polymerizable monomer capable of binding chemically to otherROMP polymerizable monomers through a ROMP chemical reaction to form apolymer. It will be understood that a polymerizable monomer may bechemically modified in the polymerization reaction to differ from thefree polymerizable monomer when forming the polymerizable monomermoiety. In embodiments, the ROMP polymerizable monomer includes anolefin. In embodiments, the ROMP polymerizable monomer includes a cyclicolefin. In embodiments, the ROMP polymerizable monomer includes a cyclicolefin with ring strain (e.g., norbornene or cyclopentene or derivativesthereof). In embodiments, the ROMP polymerizable monomer includes anoligonucleotide. In embodiments, the ROMP polymerizable monomer includesa hydrophobic moiety. In embodiments, the ROMP polymerizable monomerincludes substituted or unsubstituted norbornenyl (a monovalentsubstituted or unsubstituted norbornene). In embodiments, the ROMPpolymerizable monomer is

wherein Ring A is substituted or unsubstituted cycloalkyl, substitutedor unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R³ is independently anoligonucleotide, hydrogen, halogen, oxo, —C(halo)₃, —CH(halo)₂,—CH₂(halo), —CN, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —SH, —SO₃H, —SO₄H,—SO₂NH₂, —NHNH₂, —ONH₂, —NHC(O)NHNH₂, —NHC(O)NH₂, —NHSO₂H, —NHC(O)H,—NHC(O)OH, —NHOH, —OC(halo)₃, —OCH(halo)₂, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, or substituted or unsubstitutedheteroaryl. In embodiments, R³ is independently an oligonucleotide. Inembodiments, at least one R³ is independently an oligonucleotide. Inembodiments, R³ is independently a halogen, oxo, —C(halo)₃, —CH(halo)₂,—CH₂(halo), —CN, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —SH, —SO₃H, —SO₄H,—SO₂NH₂, —NHNH₂, —ONH₂, —NHC(O)NHNH₂, —NHC(O)NH₂, —NHSO₂H, —NHC(O)H,—NHC(O)OH, —NHOH, —OC(halo)₃, —OCH(halo)₂, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, or substituted or unsubstitutedheteroaryl. In embodiments, at least one R³ is independently anoligonucleotide. In embodiments, R³ is independently a hydrogen,halogen, oxo, —C(halo)₃, —CH(halo)₂, —CH₂(halo), —CN, —OH, —NH₂, —COOH,—CONH₂, —NO₂, —SH, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC(O)NHNH₂,—NHC(O)NH₂, —NHSO₂H, —NHC(O)H, —NHC(O)OH, —NHOH, —OC(halo)₃,—OCH(halo)₂, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl.

In embodiments, the ROMP polymerizable monomer is

In formula (IIA-IIF), Ring A, L¹, R³ and R⁴ are as defined herein. Inembodiments of formula (IIA)-(IIF), R³ is not an oligonucleotide. Inembodiments, the ROMP polymerizable monomer is

In formula (IIIA)-(IIIF), Ring A, L¹ and R4 is as defined herein.

L¹ is independently a bond, —O—, —NH—, —COO—, —S—, —SO₂—, —SO₃—, —SO₄—,—SO₂NH—, —NHC(O)—, —C(O)NH—, —NHC(O)O—, substituted or unsubstitutedalkylene, substituted or unsubstituted heteroalkylene, substituted orunsubstituted cycloalkylene, substituted or unsubstitutedheterocycloalkylene, substituted or unsubstituted arylene, orsubstituted or unsubstituted heteroarylene.

In embodiments, L¹ is independently a bond, —C(O)O—, —C(O)NH—,—C(O)NHCH₂CH₂NH—, —CH₂—, —CH₂CH₂O—, —CH₂CH₂—, —CH₂NHC(O)—,—CH₂CH₂NHC(O)—, —CH₂CH₂NH—, —CH₂O—, —CH₂CH₂N(CH₃)₂CH₂—, —CH₂C(O)—, or—CH₂CH₂C(O)—. In embodiments, L¹ is independently —C(O)O—, —C(O)NH—,—C(O)NHCH₂CH₂NH—, —CH₂—, —CH₂CH₂O—, —CH₂CH₂—, —CH₂NHC(O)—,—CH₂CH₂NHC(O)—, —CH₂CH₂NH—, —CH₂O—, —CH₂CH₂N(CH₃)₂CH₂—, —CH₂C(O)—, or—CH₂CH₂C(O)—.

R⁴ is independently an oligonucleotide as described herein.

In embodiments, a polymerizable monomer is selected from:

A “terminal polymer moiety” as used herein, refers to a chemical moietythat results from termination of a polymerization reaction (e.g., byaddition of a chain terminator or transfer agent that may be modified toform the terminal polymer moiety in the termination reaction). Terminalpolymer moieties may include a solid support, nanoparticle orappropriate termination moiety (e.g. substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl or substituted or unsubstituted heteroaryl). Inembodiments, a terminal polymer moiety includes a functional moiety, forexample a detectable moiety. In embodiments, a terminal polymer moietyis the polymerization product of an ethyl vinyl ether. In embodiments, aterminal polymer moiety is the polymerization product of an alkenecontaining substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl or substituted or unsubstituted heteroaryl. Inembodiments, a terminal polymer moiety is the polymerization product ofan alkene containing compound (e.g., also including a function group,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl,substituted or unsubstituted heteroaryl, or detectable moiety). Inembodiments, a terminal polymer moiety is selected from:

The term “ring-opening metathesis polymerization” or “ROMP” is used inaccordance with its meaning in polymer chemistry and refers to achain-growth polymerization (e.g., olefin metathesis chain-growthpolymerization). In embodiments, the reaction is driven by relief ofring strain in cyclic olegins (e.g., norbornene or cyclopentene). Inembodiments, the ROMP uses a ruthenium catalyst. In embodiments, theROMP uses a Grubbs' catalyst. In embodiments, the ROMP uses a Mocatalyst. In embodiments, the ROMP uses [Mo(═CHBut)(Nar)(OR)2]. Inembodiments, the ROMP uses a transition metal catalyst. In embodiments,the ROMP uses a transition metal carbine complex catalyst. Inembodiments, the ROMP usesBenzylidene-bis(tricyclohexylphosphine)-dichlororuthenium. Inembodiments, the ROMP uses[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(phenylmethylene)(tricyclohexylphosphine)ruthenium.In embodiments, the ROMP usesDichloro(o-isopropoxyphenylmethylene)(tricyclohexylphosphine)ruthenium(II).In embodiments, the ROMP uses[1,3-Bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(o-isopropoxyphenylmethylene)ruthenium.In embodiments, the ROMP uses a third generation Grubbs' catalyst. Inembodiments, the ROMP uses (IMesH₂)(C₅H₅N)₂(Cl)₂Ru═CHPh.

Where substituent groups are specified by their conventional chemicalformulae, written from left to right, they equally encompass thechemically identical substituents that would result from writing thestructure from right to left, e.g., —CH₂O— is equivalent to —OCH₂—.

The term “alkyl,” by itself or as part of another substituent, means,unless otherwise stated, a straight (i.e., unbranched) or branchednon-cyclic carbon chain (or carbon), or combination thereof, which maybe fully saturated, mono- or polyunsaturated and can include di- andmultivalent radicals, having the number of carbon atoms designated(i.e., C₁-C₁₀ means one to ten carbons). Examples of saturatedhydrocarbon radicals include, but are not limited to, groups such asmethyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl,sec-butyl, (cyclohexyl)methyl, homologs and isomers of, for example,n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. An unsaturated alkylgroup is one having one or more double bonds or triple bonds. Examplesof unsaturated alkyl groups include, but are not limited to, vinyl,2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl,3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and thehigher homologs and isomers. An alkoxy is an alkyl attached to theremainder of the molecule via an oxygen linker (—O—).

The term “alkylene,” by itself or as part of another substituent, means,unless otherwise stated, a divalent radical derived from an alkyl, asexemplified, but not limited by, —CH₂CH₂CH₂CH₂—. Typically, an alkyl (oralkylene) group will have from 1 to 24 carbon atoms, with those groupshaving 10 or fewer carbon atoms being preferred in the presentinvention. A “lower alkyl” or “lower alkylene” is a shorter chain alkylor alkylene group, generally having eight or fewer carbon atoms. Theterm “alkenylene,” by itself or as part of another substituent, means,unless otherwise stated, a divalent radical derived from an alkene.

The term “heteroalkyl,” by itself or in combination with another term,means, unless otherwise stated, a stable non-cyclic straight or branchedchain, or combinations thereof, including at least one carbon atom andat least one heteroatom selected from the group consisting of O, N, P,Si, and S, and wherein the nitrogen and sulfur atoms may optionally beoxidized, and the nitrogen heteroatom may optionally be quaternized. Theheteroatom(s) O, N, P, S, and Si may be placed at any interior positionof the heteroalkyl group or at the position at which the alkyl group isattached to the remainder of the molecule. Examples include, but are notlimited to: —CH₂—CH₂—O—CH₃, —CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃,—CH₂—S—CH₂—CH₃, —CH₂—CH₂, —S(O)—CH₃, —CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃,—Si(CH₃)₃, —CH₂—CH═N—OCH₃, —CH═CH—N(CH₃)—CH₃, —O—CH₃, —O—CH₂—CH₃, and—CN. Up to two or three heteroatoms may be consecutive, such as, forexample, —CH₂—NH—OCH₃ and —CH₂—O—Si(CH₃)₃.

Similarly, the term “heteroalkylene,” by itself or as part of anothersubstituent, means, unless otherwise stated, a divalent radical derivedfrom heteroalkyl, as exemplified, but not limited by,—CH₂—CH₂—S—CH₂—CH₂— and —CH₂—S—CH₂—CH₂—NH—CH₂—. For heteroalkylenegroups, heteroatoms can also occupy either or both of the chain termini(e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, andthe like). Still further, for alkylene and heteroalkylene linkinggroups, no orientation of the linking group is implied by the directionin which the formula of the linking group is written. For example, theformula —C(O)₂R′— represents both —C(O)₂R′— and —R′C(O)₂—. As describedabove, heteroalkyl groups, as used herein, include those groups that areattached to the remainder of the molecule through a heteroatom, such as—C(O)R′, —C(O)NR′, —NR′R″, —OR′, —SR′, and/or —SO₂R′. Where“heteroalkyl” is recited, followed by recitations of specificheteroalkyl groups, such as —NR′R″ or the like, it will be understoodthat the terms heteroalkyl and —NR′R″ are not redundant or mutuallyexclusive. Rather, the specific heteroalkyl groups are recited to addclarity. Thus, the term “heteroalkyl” should not be interpreted hereinas excluding specific heteroalkyl groups, such as —NR′R″ or the like.

The terms “cycloalkyl” and “heterocycloalkyl,” by themselves or incombination with other terms, mean, unless otherwise stated, cyclicnon-aromatic versions of “alkyl” and “heteroalkyl,” respectively,wherein the carbons making up the ring or rings do not necessarily needto be bonded to a hydrogen due to all carbon valencies participating inbonds with non-hydrogen atoms. Additionally, for heterocycloalkyl, aheteroatom can occupy the position at which the heterocycle is attachedto the remainder of the molecule. Examples of cycloalkyl include, butare not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples ofheterocycloalkyl include, but are not limited to,1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl,3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl,tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl,1-piperazinyl, 2-piperazinyl, and the like. A “cycloalkylene” and a“heterocycloalkylene,” alone or as part of another substituent, means adivalent radical derived from a cycloalkyl and heterocycloalkyl,respectively.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“halo(C₁-C₄)alkyl” includes, but is not limited to, fluoromethyl,difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl,3-bromopropyl, and the like.

The term “acyl” means, unless otherwise stated, —C(O)R where R is asubstituted or unsubstituted alkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl.

The term “aryl” means, unless otherwise stated, a polyunsaturated,aromatic, hydrocarbon substituent, which can be a single ring ormultiple rings (preferably from 1 to 3 rings) that are fused together(i.e., a fused ring aryl) or linked covalently (e.g., biphenyl). A fusedring aryl refers to multiple rings fused together wherein at least oneof the fused rings is an aryl ring. The term “heteroaryl” refers to arylgroups (or rings) that contain at least one heteroatom such as N, O, orS, wherein the nitrogen and sulfur atoms are optionally oxidized, andthe nitrogen atom(s) are optionally quaternized. Thus, the term“heteroaryl” includes fused ring heteroaryl groups (i.e., multiple ringsfused together wherein at least one of the fused rings is aheteroaromatic ring). A 5,6-fused ring heteroarylene refers to two ringsfused together, wherein one ring has 5 members and the other ring has 6members, and wherein at least one ring is a heteroaryl ring. Likewise, a6,6-fused ring heteroarylene refers to two rings fused together, whereinone ring has 6 members and the other ring has 6 members, and wherein atleast one ring is a heteroaryl ring. And a 6,5-fused ring heteroarylenerefers to two rings fused together, wherein one ring has 6 members andthe other ring has 5 members, and wherein at least one ring is aheteroaryl ring. A heteroaryl group can be attached to the remainder ofthe molecule through a carbon or heteroatom. Non-limiting examples ofaryl and heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl,4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl,2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl,2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl,5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl,2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl,4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl,1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl,3-quinolyl, and 6-quinolyl. Substituents for each of the above notedaryl and heteroaryl ring systems are selected from the group ofacceptable substituents described below. An “arylene” and a“heteroarylene,” alone or as part of another substituent, mean adivalent radical derived from an aryl and heteroaryl, respectively.Non-limiting examples of heteroaryl groups include pyridinyl,pyrimidinyl, thiophenyl, thienyl, furanyl, indolyl, benzoxadiazolyl,benzodioxolyl, benzodioxanyl, thianaphthanyl, pyrrolopyridinyl,indazolyl, quinolinyl, quinoxalinyl, pyridopyrazinyl, quinazolinonyl,benzoisoxazolyl, imidazopyridinyl, benzofuranyl, benzothienyl,benzothiophenyl, phenyl, naphthyl, biphenyl, pyrrolyl, pyrazolyl,imidazolyl, pyrazinyl, oxazolyl, isoxazolyl, thiazolyl, furylthienyl,pyridyl, pyrimidyl, benzothiazolyl, purinyl, benzimidazolyl,isoquinolyl, thiadiazolyl, oxadiazolyl, pyrrolyl, diazolyl, triazolyl,tetrazolyl, benzothiadiazolyl, isothiazolyl, pyrazolopyrimidinyl,pyrrolopyrimidinyl, benzotriazolyl, benzoxazolyl, or quinolyl. Theexamples above may be substituted or unsubstituted and divalent radicalsof each heteroaryl example above are non-limiting examples ofheteroarylene.

A fused ring heterocyloalkyl-aryl is an aryl fused to aheterocycloalkyl. A fused ring heterocycloalkyl-heteroaryl is aheteroaryl fused to a heterocycloalkyl. A fused ringheterocycloalkyl-cycloalkyl is a heterocycloalkyl fused to a cycloalkyl.A fused ring heterocycloalkyl-heterocycloalkyl is a heterocycloalkylfused to another heterocycloalkyl. Fused ring heterocycloalkyl-aryl,fused ring heterocycloalkyl-heteroaryl, fused ringheterocycloalkyl-cycloalkyl, or fused ringheterocycloalkyl-heterocycloalkyl may each independently beunsubstituted or substituted with one or more of the substitutentsdescribed herein.

The term “oxo,” as used herein, means an oxygen that is double bonded toa carbon atom.

The term “alkylsulfonyl,” as used herein, means a moiety having theformula —S(O₂)—R′, where R′ is a substituted or unsubstituted alkylgroup as defined above. R′ may have a specified number of carbons (e.g.,“C₁-C₄ alkylsulfonyl”).

Each of the above terms (e.g., “alkyl,” “heteroalkyl,” “aryl,” and“heteroaryl”) includes both substituted and unsubstituted forms of theindicated radical. Preferred substituents for each type of radical areprovided below.

Substituents for the alkyl and heteroalkyl radicals (including thosegroups often referred to as alkylene, alkenyl, heteroalkylene,heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, andheterocycloalkenyl) can be one or more of a variety of groups selectedfrom, but not limited to, —OR′, ═O, ═NR′,

═N—OR′, —NR′R″, —SR′, -halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′,—CONR′R″, —OC(O)N R′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′,—NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″,—NRSO₂R′, —NR′NR″R′″, —ONR′R″, —NR′C═(O)NR″NR″′R″″, —CN, —NO₂, in anumber ranging from zero to (2m′+1), where m′ is the total number ofcarbon atoms in such radical. R, R′, R″, R′″, and R″″ each preferablyindependently refer to hydrogen, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl (e.g.,aryl substituted with 1-3 halogens), substituted or unsubstitutedheteroaryl, substituted or unsubstituted alkyl, alkoxy, or thioalkoxygroups, or arylalkyl groups. When a compound of the invention includesmore than one R group, for example, each of the R groups isindependently selected as are each R′, R″, R′″, and R″″ group when morethan one of these groups is present. When R′ and R″ are attached to thesame nitrogen atom, they can be combined with the nitrogen atom to forma 4-, 5-, 6-, or 7-membered ring. For example, —NR′R″ includes, but isnot limited to, 1-pyrrolidinyl and 4-morpholinyl. From the abovediscussion of substituents, one of skill in the art will understand thatthe term “alkyl” is meant to include groups including carbon atoms boundto groups other than hydrogen groups, such as haloalkyl (e.g., —CF₃ and—CH₂CF₃) and acyl (e.g., —C(O)CH₃, —C(O)CF₃, —C(O)CH₂OCH₃, and thelike).

Similar to the substituents described for the alkyl radical,substituents for the aryl and heteroaryl groups are varied and areselected from, for example: —OR′, —NR′R″, —SR′, -halogen, —SiR′R″R′″,—OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC (O)NR′R″, —NR″C(O)R′,—NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″,—S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NRSO₂R′, —NR′NR″R′″, —ONR′R″,—NR′C═(O)NR″NR″′R″″, —CN, —NO₂, —R′, —N₃, —CH(Ph)₂, fluoro(C₁-C₄)alkoxy,and fluoro(C₁-C₄)alkyl, in a number ranging from zero to the totalnumber of open valences on the aromatic ring system; and where R′, R″,R′″, and R″″ are preferably independently selected from hydrogen,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. When a compound of theinvention includes more than one R group, for example, each of the Rgroups is independently selected as are each R′, R″, R′″, and R″″ groupswhen more than one of these groups is present.

Two or more substituents may optionally be joined to form aryl,heteroaryl, cycloalkyl, or heterocycloalkyl groups. Such so-calledring-forming substituents are typically, though not necessarily, foundattached to a cyclic base structure. In one embodiment, the ring-formingsubstituents are attached to adjacent members of the base structure. Forexample, two ring-forming substituents attached to adjacent members of acyclic base structure create a fused ring structure. In anotherembodiment, the ring-forming substituents are attached to a singlemember of the base structure. For example, two ring-forming substituentsattached to a single member of a cyclic base structure create aspirocyclic structure. In yet another embodiment, the ring-formingsubstituents are attached to non-adjacent members of the base structure.

Two of the substituents on adjacent atoms of the aryl or heteroaryl ringmay optionally form a ring of the formula -T-C(O)—(CRR′)_(q)—U—, whereinT and U are independently —NR—, —O—, —CRR′—, or a single bond, and q isan integer of from 0 to 3. Alternatively, two of the substituents onadjacent atoms of the aryl or heteroaryl ring may optionally be replacedwith a substituent of the formula -A-(CH₂)_(r)—B—, wherein A and B areindependently —CRR′—, —O—, —NR—, —S—, —S(O)—, —S(O)₂—, —S(O)₂NR′—, or asingle bond, and r is an integer of from 1 to 4. One of the single bondsof the new ring so formed may optionally be replaced with a double bond.Alternatively, two of the substituents on adjacent atoms of the aryl orheteroaryl ring may optionally be replaced with a substituent of theformula —(CRR′)_(s)—X′— (C″R″R′″)_(d)—, where s and d are independentlyintegers of from 0 to 3, and X′ is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or—S(O)₂NR′—. The substituents R, R′, R″, and R′″ are preferablyindependently selected from hydrogen, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, and substituted or unsubstitutedheteroaryl.

As used herein, the terms “heteroatom” or “ring heteroatom” are meant toinclude, oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), andsilicon (Si).

A “substituent group,” as used herein, means a group selected from thefollowing moieties:

-   -   (A) oxo, halogen, —CF₃, —CN, —OH, —NH₂, —COOH, —CONH₂, —NO₂,        —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,        —NHC═(O)NHNH₂, —NHC═(O) NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH,        —NHOH, —OCF₃, —OCHF₂, —NHSO₂CH₃, —N₃, unsubstituted alkyl,        unsubstituted heteroalkyl, unsubstituted cycloalkyl,        unsubstituted heterocycloalkyl, unsubstituted aryl,        unsubstituted heteroaryl, and    -   (B) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,        heteroaryl, substituted with at least one substituent selected        from:        -   (i) oxo, halogen, —CF₃, —CN, —OH, —NH₂, —COOH, —CONH₂, —NO₂,            —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,            —NHC═(O)NHNH₂, —NHC═(O) NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH,            —NHOH, —OCF₃, —OCHF₂, —NHSO₂CH₃, —N₃, unsubstituted alkyl,            unsubstituted heteroalkyl, unsubstituted cycloalkyl,            unsubstituted heterocycloalkyl, unsubstituted aryl,            unsubstituted heteroaryl, and        -   (ii) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,            heteroaryl, substituted with at least one substituent            selected from:            -   (a) oxo, halogen, —CF₃, —CN, —OH, —NH₂, —COOH, —CONH₂,                —NO₂, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,                —NHC═(O)NHNH₂, —NHC═(O) NH₂, —NHSO₂H, —NHC═(O)H,                —NHC(O)—OH, —NHOH, —OCF₃, —OCHF₂, —NHSO₂CH₃, —N₃,                unsubstituted alkyl, unsubstituted heteroalkyl,                unsubstituted cycloalkyl, unsubstituted                heterocycloalkyl, unsubstituted aryl, unsubstituted                heteroaryl, and            -   (b) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,                aryl, heteroaryl, substituted with at least one                substituent selected from: oxo,            -   halogen, —CF₃, —CN, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —SH,                —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,                —NHC═(O)NHNH₂, —NHC═(O) NH₂, —NHSO₂H, —NHC═(O)H,                —NHC(O)—OH, —NHOH, —OCF₃, —OCHF₂, —NHSO₂CH₃, —N₃,                unsubstituted alkyl, unsubstituted heteroalkyl,                unsubstituted cycloalkyl, unsubstituted                heterocycloalkyl, unsubstituted aryl, unsubstituted                heteroaryl.

A “size-limited substituent” or “size-limited substituent group,” asused herein, means a group selected from all of the substituentsdescribed above for a “substituent group,” wherein each substituted orunsubstituted alkyl is a substituted or unsubstituted C₁-C₂₀ alkyl, eachsubstituted or unsubstituted heteroalkyl is a substituted orunsubstituted 2 to 20 membered heteroalkyl, each substituted orunsubstituted cycloalkyl is a substituted or unsubstituted C₃-C₈cycloalkyl, each substituted or unsubstituted heterocycloalkyl is asubstituted or unsubstituted 3 to 8 membered heterocycloalkyl, eachsubstituted or unsubstituted aryl is a substituted or unsubstitutedC₆-C₁₀ aryl, and each substituted or unsubstituted heteroaryl is asubstituted or unsubstituted 5 to 10 membered heteroaryl.

A “lower substituent” or “lower substituent group,” as used herein,means a group selected from all of the substituents described above fora “substituent group,” wherein each substituted or unsubstituted alkylis a substituted or unsubstituted C₁-C₈ alkyl, each substituted orunsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8membered heteroalkyl, each substituted or unsubstituted cycloalkyl is asubstituted or unsubstituted C₃-C₇ cycloalkyl, each substituted orunsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 7membered heterocycloalkyl, each substituted or unsubstituted aryl is asubstituted or unsubstituted C₆-C₁₀ aryl, and each substituted orunsubstituted heteroaryl is a substituted or unsubstituted 5 to 9membered heteroaryl.

In some embodiments, each substituted group described in the compoundsherein is substituted with at least one substituent group. Morespecifically, in some embodiments, each substituted alkyl, substitutedheteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl,substituted aryl, substituted heteroaryl, substituted alkylene,substituted heteroalkylene, substituted cycloalkylene, substitutedheterocycloalkylene, substituted arylene, and/or substitutedheteroarylene described in the compounds herein are substituted with atleast one substituent group. In other embodiments, at least one or allof these groups are substituted with at least one size-limitedsubstituent group. In other embodiments, at least one or all of thesegroups are substituted with at least one lower substituent group.

In other embodiments of the compounds herein, each substituted orunsubstituted alkyl may be a substituted or unsubstituted C₁-C₂₀ alkyl,each substituted or unsubstituted heteroalkyl is a substituted orunsubstituted 2 to 20 membered heteroalkyl, each substituted orunsubstituted cycloalkyl is a substituted or unsubstituted C₃-C₈cycloalkyl, each substituted or unsubstituted heterocycloalkyl is asubstituted or unsubstituted 3 to 8 membered heterocycloalkyl, eachsubstituted or unsubstituted aryl is a substituted or unsubstitutedC₆-C₁₀ aryl, and/or each substituted or unsubstituted heteroaryl is asubstituted or unsubstituted 5 to 10 membered heteroaryl. In someembodiments of the compounds herein, each substituted or unsubstitutedalkylene is a substituted or unsubstituted C₁-C₂₀ alkylene, eachsubstituted or unsubstituted heteroalkylene is a substituted orunsubstituted 2 to 20 membered heteroalkylene, each substituted orunsubstituted cycloalkylene is a substituted or unsubstituted C₃-C₈cycloalkylene, each substituted or unsubstituted heterocycloalkylene isa substituted or unsubstituted 3 to 8 membered heterocycloalkylene, eachsubstituted or unsubstituted arylene is a substituted or unsubstitutedC₆-C₁₀ arylene, and/or each substituted or unsubstituted heteroaryleneis a substituted or unsubstituted 5 to 10 membered heteroarylene.

In some embodiments, each substituted or unsubstituted alkyl is asubstituted or unsubstituted C₁-C₈ alkyl, each substituted orunsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8membered heteroalkyl, each substituted or unsubstituted cycloalkyl is asubstituted or unsubstituted C₃-C₇ cycloalkyl, each substituted orunsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 7membered heterocycloalkyl, each substituted or unsubstituted aryl is asubstituted or unsubstituted C₆-C₁₀ aryl, and/or each substituted orunsubstituted heteroaryl is a substituted or unsubstituted 5 to 9membered heteroaryl. In some embodiments, each substituted orunsubstituted alkylene is a substituted or unsubstituted C₁-C₈ alkylene,each substituted or unsubstituted heteroalkylene is a substituted orunsubstituted 2 to 8 membered heteroalkylene, each substituted orunsubstituted cycloalkylene is a substituted or unsubstituted C₃-C₇cycloalkylene, each substituted or unsubstituted heterocycloalkylene isa substituted or unsubstituted 3 to 7 membered heterocycloalkylene, eachsubstituted or unsubstituted arylene is a substituted or unsubstitutedC₆-C₁₀ arylene, and/or each substituted or unsubstituted heteroaryleneis a substituted or unsubstituted 5 to 9 membered heteroarylene. In someembodiments, the compound is a chemical species set forth in theExamples section below.

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds that are prepared with relatively nontoxic acidsor bases, depending on the particular substituents found on thecompounds described herein. When compounds of the present inventioncontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable base additionsalts include sodium, potassium, calcium, ammonium, organic amino, ormagnesium salt, or a similar salt. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, maleic, malonic, benzoic, succinic,suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic,p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Alsoincluded are salts of amino acids such as arginate and the like, andsalts of organic acids like glucuronic or galactunoric acids and thelike (see, e.g., Berge et al., Journal of Pharmaceutical Science 66:1-19(1977)). Certain specific compounds of the present invention containboth basic and acidic functionalities that allow the compounds to beconverted into either base or acid addition salts. Otherpharmaceutically acceptable carriers known to those of skill in the artare suitable for the present invention. Salts tend to be more soluble inaqueous or other protonic solvents that are the corresponding free baseforms. In other cases, the preparation may be a lyophilized powder in 1mM-50 mM histidine, 0.1%-2% sucrose, 2%-7% mannitol at a pH range of 4.5to 5.5, that is combined with buffer prior to use.

Thus, the compounds of the present invention may exist as salts, such aswith pharmaceutically acceptable acids. The present invention includessuch salts. Examples of such salts include hydrochlorides,hydrobromides, sulfates, methanesulfonates, nitrates, maleates,acetates, citrates, fumarates, tartrates (e.g., (+)-tartrates,(−)-tartrates, or mixtures thereof including racemic mixtures),succinates, benzoates, and salts with amino acids such as glutamic acid.These salts may be prepared by methods known to those skilled in theart.

The neutral forms of the compounds are preferably regenerated bycontacting the salt with a base or acid and isolating the parentcompound in the conventional manner. The parent form of the compounddiffers from the various salt forms in certain physical properties, suchas solubility in polar solvents.

In addition to salt forms, the present invention provides compounds,which are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are encompassedwithin the scope of the present invention. Certain compounds of thepresent invention may exist in multiple crystalline or amorphous forms.In general, all physical forms are equivalent for the uses contemplatedby the present invention and are intended to be within the scope of thepresent invention.

As used herein, the term “salt” refers to acid or base salts of thecompounds used in the methods of the present invention. Illustrativeexamples of acceptable salts are mineral acid (hydrochloric acid,hydrobromic acid, phosphoric acid, and the like) salts, organic acid(acetic acid, propionic acid, glutamic acid, citric acid and the like)salts, quaternary ammonium (methyl iodide, ethyl iodide, and the like)salts.

Certain compounds of the present invention possess asymmetric carbonatoms (optical or chiral centers) or double bonds; the enantiomers,racemates, diastereomers, tautomers, geometric isomers, stereoisometricforms that may be defined, in terms of absolute stereochemistry, as (R)-or (S)- or, as (D)- or (L)- for amino acids, and individual isomers areencompassed within the scope of the present invention. The compounds ofthe present invention do not include those which are known in art to betoo unstable to synthesize and/or isolate. The present invention ismeant to include compounds in racemic and optically pure forms.Optically active (R)- and (S)-, or (D)- and (L)-isomers may be preparedusing chiral synthons or chiral reagents, or resolved using conventionaltechniques. When the compounds described herein contain olefinic bondsor other centers of geometric asymmetry, and unless specified otherwise,it is intended that the compounds include both E and Z geometricisomers.

As used herein, the term “isomers” refers to compounds having the samenumber and kind of atoms, and hence the same molecular weight, butdiffering in respect to the structural arrangement or configuration ofthe atoms.

The term “tautomer,” as used herein, refers to one of two or morestructural isomers which exist in equilibrium and which are readilyconverted from one isomeric form to another.

It will be apparent to one skilled in the art that certain compounds ofthis invention may exist in tautomeric forms, all such tautomeric formsof the compounds being within the scope of the invention.

Unless otherwise stated, structures depicted herein are also meant toinclude all stereochemical forms of the structure; i.e., the R and Sconfigurations for each asymmetric center. Therefore, singlestereochemical isomers as well as enantiomeric and diastereomericmixtures of the present compounds are within the scope of the invention.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds which differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement of a hydrogen by a deuterium ortritium, or the replacement of a carbon by ¹³C- or ¹⁴C-enriched carbonare within the scope of this invention.

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine-125 (¹²⁵I), or carbon-14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areencompassed within the scope of the present invention.

The symbol “

” denotes the point of attachment of a chemical moiety to the remainderof a molecule or chemical formula.

The terms “a” or “an,” as used in herein means one or more. In addition,the phrase “substituted with a[n],” as used herein, means the specifiedgroup may be substituted with one or more of any or all of the namedsubstituents. For example, where a group, such as an alkyl or heteroarylgroup, is “substituted with an unsubstituted C₁-C₂₀ alkyl, orunsubstituted 2 to 20 membered heteroalkyl,” the group may contain oneor more unsubstituted C₁-C₂₀ alkyls, and/or one or more unsubstituted 2to 20 membered heteroalkyls. Moreover, where a moiety is substitutedwith an R substituent, the group may be referred to as “R-substituted.”Where a moiety is R-substituted, the moiety is substituted with at leastone R substituent and each R substituent is optionally different.

Descriptions of compounds of the present invention are limited byprinciples of chemical bonding known to those skilled in the art.Accordingly, where a group may be substituted by one or more of a numberof substituents, such substitutions are selected so as to comply withprinciples of chemical bonding and to give compounds which are notinherently unstable and/or would be known to one of ordinary skill inthe art as likely to be unstable under ambient conditions, such asaqueous, neutral, and several known physiological conditions. Forexample, a heterocycloalkyl or heteroaryl is attached to the remainderof the molecule via a ring heteroatom in compliance with principles ofchemical bonding known to those skilled in the art thereby avoidinginherently unstable compounds.

The terms “treating” or “treatment” refers to any indicia of success inthe treatment or amelioration of an injury, disease, pathology orcondition, including any objective or subjective parameter such asabatement; remission; diminishing of symptoms or making the injury,pathology or condition more tolerable to the patient; slowing in therate of degeneration or decline; making the final point of degenerationless debilitating; improving a patient's physical or mental well-being.The treatment or amelioration of symptoms can be based on objective orsubjective parameters; including the results of a physical examination,neuropsychiatric exams, and/or a psychiatric evaluation.

An “effective amount” is an amount sufficient to accomplish a statedpurpose (e.g. achieve the effect for which it is administered, treat adisease, reduce enzyme activity, increase enzyme activity, reducetranscriptional activity, increase transcriptional activity, reduce oneor more symptoms of a disease or condition). An example of an “effectiveamount” is an amount sufficient to contribute to the treatment,prevention, or reduction of a symptom or symptoms of a disease, whichcould also be referred to as a “therapeutically effective amount.” A“reduction” of a symptom or symptoms (and grammatical equivalents ofthis phrase) means decreasing of the severity or frequency of thesymptom(s), or elimination of the symptom(s). A “prophylacticallyeffective amount” of a drug is an amount of a drug that, whenadministered to a subject, will have the intended prophylactic effect,e.g., preventing or delaying the onset (or reoccurrence) of an injury,disease, pathology or condition, or reducing the likelihood of the onset(or reoccurrence) of an injury, disease, pathology, or condition, ortheir symptoms. The full prophylactic effect does not necessarily occurby administration of one dose, and may occur only after administrationof a series of doses. Thus, a prophylactically effective amount may beadministered in one or more administrations. An “activity decreasingamount,” as used herein, refers to an amount of antagonist (inhibitor)required to decrease the activity of an enzyme or protein (e.g.transcription factor) relative to the absence of the antagonist. An“activity increasing amount,” as used herein, refers to an amount ofagonist (activator) required to increase the activity of an enzyme orprotein (e.g. transcription factor) relative to the absence of theagonist. A “function disrupting amount,” as used herein, refers to theamount of antagonist (inhibitor) required to disrupt the function of anenzyme or protein (e.g. transcription factor) relative to the absence ofthe antagonist. A “function increasing amount,” as used herein, refersto the amount of agonist (activator) required to increase the functionof an enzyme or protein (e.g. transcription factor) relative to theabsence of the agonist. The exact amounts will depend on the purpose ofthe treatment, and will be ascertainable by one skilled in the art usingknown techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms(vols. 1-3, 1992); Lloyd, The Art, Science and Technology ofPharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999);and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003,Gennaro, Ed., Lippincott, Williams & Wilkins).

The term “associated” or “associated with” in the context of a substanceor substance activity or function associated with a disease means thatthe disease is caused by (in whole or in part), or a symptom of thedisease is caused by (in whole or in part) the substance or substanceactivity or function.

“Control” or “control experiment” is used in accordance with its plainordinary meaning and refers to an experiment in which the subjects orreagents of the experiment are treated as in a parallel experimentexcept for omission of a procedure, reagent, or variable of theexperiment. In some instances, the control is used as a standard ofcomparison in evaluating experimental effects.

“Contacting” is used in accordance with its plain ordinary meaning andrefers to the process of allowing at least two distinct species (e.g.chemical compounds including biomolecules, or cells) to becomesufficiently proximal to react, interact or physically touch. It shouldbe appreciated, however, that the resulting reaction product can beproduced directly from a reaction between the added reagents or from anintermediate from one or more of the added reagents which can beproduced in the reaction mixture. The term “contacting” may includeallowing two species to react, interact, or physically touch, whereinthe two species may be a compound as described herein and a protein orenzyme. In some embodiments contacting includes allowing a compounddescribed herein to interact with a protein or enzyme that is involvedin a signaling pathway.

As defined herein, the term “inhibition”, “inhibit”, “inhibiting” andthe like in reference to a protein-inhibitor (e.g. antagonist)interaction means negatively affecting (e.g. decreasing) the activity orfunction of the protein relative to the activity or function of theprotein in the absence of the inhibitor. In some embodiments inhibitionrefers to reduction of a disease or symptoms of disease. In someembodiments, inhibition refers to a reduction in the activity of asignal transduction pathway or signaling pathway. Thus, inhibitionincludes, at least in part, partially or totally blocking stimulation,decreasing, preventing, or delaying activation, or inactivating,desensitizing, or down-regulating signal transduction or enzymaticactivity or the amount of a protein.

As defined herein, the term “activation”, “activate”, “activating” andthe like in reference to a protein-activator (e.g. agonist) interactionmeans positively affecting (e.g. increasing) the activity or function ofthe protein

The term “modulator” refers to a composition that increases or decreasesthe level of a target molecule or the function of a target molecule.

“Patient” or “subject in need thereof” refers to a living organismsuffering from or prone to a disease or condition that can be treated byadministration of a compound or pharmaceutical composition, as providedherein. Non-limiting examples include humans, other mammals, bovines,rats, mice, dogs, monkeys, goat, sheep, cows, deer, and othernon-mammalian animals. In some embodiments, a patient is human. In someembodiments, a patient is a mammal. In some embodiments, a patient is amouse. In some embodiments, a patient is an experimental animal. In someembodiments, a patient is a rat. In some embodiments, a patient is atest animal.

“Disease” or “condition” refer to a state of being or health status of apatient or subject capable of being treated with a compound,pharmaceutical composition, or method provided herein.

“Pharmaceutically acceptable excipient” and “pharmaceutically acceptablecarrier” refer to a substance that aids the administration of an activeagent to and absorption by a subject and can be included in thecompositions of the present invention without causing a significantadverse toxicological effect on the patient. Non-limiting examples ofpharmaceutically acceptable excipients include water, NaCl, normalsaline solutions, lactated Ringer's, normal sucrose, normal glucose,binders, fillers, disintegrants, lubricants, coatings, sweeteners,flavors, salt solutions (such as Ringer's solution), alcohols, oils,gelatins, carbohydrates such as lactose, amylose or starch, fatty acidesters, hydroxymethycellulose, polyvinyl pyrrolidine, and colors, andthe like. Such preparations can be sterilized and, if desired, mixedwith auxiliary agents such as lubricants, preservatives, stabilizers,wetting agents, emulsifiers, salts for influencing osmotic pressure,buffers, coloring, and/or aromatic substances and the like that do notdeleteriously react with the compounds of the invention. One of skill inthe art will recognize that other pharmaceutical excipients are usefulin the present invention.

The term “preparation” is intended to include the formulation of theactive compound with encapsulating material as a carrier providing acapsule in which the active component with or without other carriers, issurrounded by a carrier, which is thus in association with it.Similarly, cachets and lozenges are included. Tablets, powders,capsules, pills, cachets, and lozenges can be used as solid dosage formssuitable for oral administration.

As used herein, the term “administering” means oral administration,administration as a suppository, topical contact, intravenous,parenteral, intraperitoneal, intramuscular, intralesional, intrathecal,intracranial, intranasal or subcutaneous administration, or theimplantation of a slow-release device, e.g., a mini-osmotic pump, to asubject. Administration is by any route, including parenteral andtransmucosal (e.g., buccal, sublingual, palatal, gingival, nasal,vaginal, rectal, or transdermal). In embodiments, administrationincludes direct administration to a tumor. Parenteral administrationincludes, e.g., intravenous, intramuscular, intra-arteriole,intradermal, subcutaneous, intraperitoneal, intraventricular, andintracranial. Other modes of delivery include, but are not limited to,the use of liposomal formulations, intravenous infusion, transdermalpatches, etc. By “co-administer” it is meant that a compositiondescribed herein is administered at the same time, just prior to, orjust after the administration of one or more additional therapies (e.g.anti-cancer agent or chemotherapeutic). The compound of the inventioncan be administered alone or can be coadministered to the patient.Coadministration is meant to include simultaneous or sequentialadministration of the compound individually or in combination (more thanone compound or agent). Thus, the preparations can also be combined,when desired, with other active substances (e.g. to reduce metabolicdegradation). The compositions of the present invention can be deliveredby transdermally, by a topical route, formulated as applicator sticks,solutions, suspensions, emulsions, gels, creams, ointments, pastes,jellies, paints, powders, and aerosols. Oral preparations includetablets, pills, powder, dragees, capsules, liquids, lozenges, cachets,gels, syrups, slurries, suspensions, etc., suitable for ingestion by thepatient. Solid form preparations include powders, tablets, pills,capsules, cachets, suppositories, and dispersible granules. Liquid formpreparations include solutions, suspensions, and emulsions, for example,water or water/propylene glycol solutions. The compositions of thepresent invention may additionally include components to providesustained release and/or comfort. Such components include high molecularweight, anionic mucomimetic polymers, gelling polysaccharides andfinely-divided drug carrier substrates. These components are discussedin greater detail in U.S. Pat. Nos. 4,911,920; 5,403,841; 5,212,162; and4,861,760. The entire contents of these patents are incorporated hereinby reference in their entirety for all purposes. The compositions of thepresent invention can also be delivered as microspheres for slow releasein the body. For example, microspheres can be administered viaintradermal injection of drug-containing microspheres, which slowlyrelease subcutaneously (see Rao, J. Biomater Sci. Polym. Ed. 7:623-645,1995; as biodegradable and injectable gel formulations (see, e.g., GaoPharm. Res. 12:857-863, 1995); or, as microspheres for oraladministration (see, e.g., Eyles, J. Pharm. Pharmacol. 49:669-674,1997). In another embodiment, the formulations of the compositions ofthe present invention can be delivered by the use of liposomes whichfuse with the cellular membrane or are endocytosed, i.e., by employingreceptor ligands attached to the liposome, that bind to surface membraneprotein receptors of the cell resulting in endocytosis. By usingliposomes, particularly where the liposome surface carries receptorligands specific for target cells, or are otherwise preferentiallydirected to a specific organ, one can focus the delivery of thecompositions of the present invention into the target cells in vivo.(See, e.g., Al-Muhammed, J. Microencapsul. 13:293-306, 1996; Chonn,Curr. Opin. Biotechnol. 6:698-708, 1995; Ostro, Am. J. Hosp. Pharm.46:1576-1587, 1989). The compositions of the present invention can alsobe delivered as nanoparticles.

“Nucleic acid” or “oligonucleotide” or “polynucleotide” or grammaticalequivalents used herein means at least two nucleotides covalently linkedtogether. The term “nucleic acid” includes single-, double-, ormultiple-stranded DNA, RNA and analogs (derivatives) thereof.Oligonucleotides are typically from about 5, 6, 7, 8, 9, 10, 12, 15, 25,30, 40, 50 or more nucleotides in length, up to about 100 nucleotides inlength. Nucleic acids and polynucleotides are a polymers of any length,including longer lengths, e.g., 200, 300, 500, 1000, 2000, 3000, 5000,7000, 10,000, etc. In certain embodiments. the nucleic acids hereincontain phosphodiester bonds. In other embodiments, nucleic acid analogsare included that may have alternate backbones (e.g. phosphodiesterderivatives), including, e.g., phosphoramidate, phosphorodiamidate,phosphorothioate (also known as phosphothioate), phosphorodithioate,phosphonocarboxylic acids, phosphonocarboxylates, phosphonoacetic acid,phosphonoformic acid, methyl phosphonate, boron phosphonate, peptidenucleic acid linkages, or O-methylphosphoroamidite linkages (seeEckstein, Oligonucleotides and Analogues: A Practical Approach, OxfordUniversity Press), and peptide nucleic acid backbones and linkages.Other analog nucleic acids include those with positive backbones;non-ionic backbones, modified sugars, and non-ribose backbones (e.g.phosphorodiamidate morpholino oligos or locked nucleic acids (LNA)),including those described in U.S. Pat. Nos. 5,235,033 and 5,034,506, andChapters 6 and 7, ASC Symposium Series 580, Carbohydrate Modificationsin Antisense Research, Sanghui & Cook, eds. Nucleic acids containing oneor more carbocyclic sugars are also included within one definition ofnucleic acids. Modifications of the ribose-phosphate backbone may bedone for a variety of reasons, e.g., to increase the stability andhalf-life of such molecules in physiological environments or as probeson a biochip. Mixtures of naturally occurring nucleic acids and analogscan be made; alternatively, mixtures of different nucleic acid analogs,and mixtures of naturally occurring nucleic acids and analogs may bemade. In embodiments, the internucleotide linkages in DNA arephosphodiester, phosphodiester derivatives, or a combination of both.

A particular nucleic acid sequence also encompasses “splice variants.”Similarly, a particular protein encoded by a nucleic acid encompassesany protein encoded by a splice variant of that nucleic acid. “Splicevariants,” as the name suggests, are products of alternative splicing ofa gene. After transcription, an initial nucleic acid transcript may bespliced such that different (alternate) nucleic acid splice productsencode different polypeptides. Mechanisms for the production of splicevariants vary, but include alternate splicing of exons. Alternatepolypeptides derived from the same nucleic acid by read-throughtranscription are also encompassed by this definition. Any products of asplicing reaction, including recombinant forms of the splice products,are included in this definition. An example of potassium channel splicevariants is discussed in Leicher, et al., J. Biol. Chem.273(52):35095-35101 (1998).

Nucleic acid is “operably linked” when it is placed into a functionalrelationship with another nucleic acid sequence. For example, DNA for apresequence or secretory leader is operably linked to DNA for apolypeptide if it is expressed as a preprotein that participates in thesecretion of the polypeptide; a promoter or enhancer is operably linkedto a coding sequence if it affects the transcription of the sequence; ora ribosome binding site is operably linked to a coding sequence if it ispositioned so as to facilitate translation. Generally, “operably linked”means that the DNA sequences being linked are near each other, and, inthe case of a secretory leader, contiguous and in reading phase.However, enhancers do not have to be contiguous. Linking is accomplishedby ligation at convenient restriction sites. If such sites do not exist,the synthetic oligonucleotide adaptors or linkers are used in accordancewith conventional practice.

An amino acid residue in a protein “corresponds” to a given residue whenit occupies the same essential structural position within the protein asthe given residue.

The terms “identical” or percent “identity,” in the context of two ormore nucleic acids or polypeptide sequences, refer to two or moresequences or subsequences that are the same or have a specifiedpercentage of amino acid residues or nucleotides that are the same(i.e., about 60% identity, preferably 61%, 62%, 63%, 64%, 65%, 66%, 67%,68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%,82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99% or higher identity over a specified region whencompared and aligned for maximum correspondence over a comparison windowor designated region) as measured using a BLAST or BLAST 2.0 sequencecomparison algorithms with default parameters described below, or bymanual alignment and visual inspection (see, e.g., NCBI web site or thelike). Such sequences are then said to be “substantially identical.”This definition also refers to, or may be applied to, the compliment ofa test sequence. The definition also includes sequences that havedeletions and/or additions, as well as those that have substitutions. Asdescribed below, the preferred algorithms can account for gaps and thelike. Preferably, identity exists over a region that is at least about10 amino acids or 20 nucleotides in length, or more preferably over aregion that is 10-50 amino acids or 20-50 nucleotides in length. As usedherein, percent (%) amino acid sequence identity is defined as thepercentage of amino acids in a candidate sequence that are identical tothe amino acids in a reference sequence, after aligning the sequencesand introducing gaps, if necessary, to achieve the maximum percentsequence identity. Alignment for purposes of determining percentsequence identity can be achieved in various ways that are within theskill in the art, for instance, using publicly available computersoftware such as BLAST, BLAST-2, ALIGN, ALIGN-2 or Megalign (DNASTAR)software. Appropriate parameters for measuring alignment, including anyalgorithms needed to achieve maximal alignment over the full-length ofthe sequences being compared can be determined by known methods.

For sequence comparisons, typically one sequence acts as a referencesequence, to which test sequences are compared. When using a sequencecomparison algorithm, test and reference sequences are entered into acomputer, subsequence coordinates are designated, if necessary, andsequence algorithm program parameters are designated. Preferably,default program parameters can be used, or alternative parameters can bedesignated. The sequence comparison algorithm then calculates thepercent sequence identities for the test sequences relative to thereference sequence, based on the program parameters.

A “comparison window”, as used herein, includes reference to a segmentof any one of the number of contiguous positions selected from the groupconsisting of from 10 to 600, usually about 50 to about 200, moreusually about 100 to about 150 in which a sequence may be compared to areference sequence of the same number of contiguous positions after thetwo sequences are optimally aligned. Methods of alignment of sequencesfor comparison are well-known in the art. Optimal alignment of sequencesfor comparison can be conducted, e.g., by the local homology algorithmof Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homologyalignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970),by the search for similarity method of Pearson & Lipman, Proc. Nat'l.Acad. Sci. USA 85:2444 (1988), by computerized implementations of thesealgorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin GeneticsSoftware Package, Genetics Computer Group, 575 Science Dr., Madison,Wis.), or by manual alignment and visual inspection (see, e.g., CurrentProtocols in Molecular Biology (Ausubel et al., eds. 1995 supplement)).

The phrase “selectively (or specifically) hybridizes to” refers to thebinding, duplexing, or hybridizing of a molecule only to a particularnucleotide sequence with a higher affinity, e.g., under more stringentconditions, than to other nucleotide sequences (e.g., total cellular orlibrary DNA or RNA).

The phrase “stringent hybridization conditions” refers to conditionsunder which a probe will hybridize to its target subsequence, typicallyin a complex mixture of nucleic acids, but to no other sequences.Stringent conditions are sequence-dependent and will be different indifferent circumstances. Longer sequences hybridize specifically athigher temperatures. An extensive guide to the hybridization of nucleicacids is found in Tijssen, Techniques in Biochemistry and MolecularBiology—Hybridization with Nucleic Probes, “Overview of principles ofhybridization and the strategy of nucleic acid assays” (1993).Generally, stringent conditions are selected to be about 5-10° C. lowerthan the thermal melting point (T_(m)) for the specific sequence at adefined ionic strength pH. The T_(m) is the temperature (under definedionic strength, pH, and nucleic concentration) at which 50% of theprobes complementary to the target hybridize to the target sequence atequilibrium (as the target sequences are present in excess, at T_(m),50% of the probes are occupied at equilibrium). Stringent conditions mayalso be achieved with the addition of destabilizing agents such asformamide. For selective or specific hybridization, a positive signal isat least two times background, preferably 10 times backgroundhybridization. Exemplary stringent hybridization conditions can be asfollowing: 50% formamide, 5×SSC, and 1% SDS, incubating at 42° C., or,5×SSC, 1% SDS, incubating at 65° C., with wash in 0.2×SSC, and 0.1% SDSat 65° C.

Nucleic acids that do not hybridize to each other under stringentconditions are still substantially identical if the polypeptides whichthey encode are substantially identical. This occurs, for example, whena copy of a nucleic acid is created using the maximum codon degeneracypermitted by the genetic code. In such cases, the nucleic acidstypically hybridize under moderately stringent hybridization conditions.Exemplary “moderately stringent hybridization conditions” include ahybridization in a buffer of 40% formamide, 1 M NaCl, 1% SDS at 37° C.,and a wash in 1×SSC at 45° C. A positive hybridization is at least twicebackground. Those of ordinary skill will readily recognize thatalternative hybridization and wash conditions can be utilized to provideconditions of similar stringency. Additional guidelines for determininghybridization parameters are provided in numerous reference, e.g., andCurrent Protocols in Molecular Biology, ed. Ausubel, et al.

A “label” or a “detectable moiety” is a composition detectable byspectroscopic, photochemical, biochemical, immunochemical, chemical,magnetic resonance imaging, or other physical means. For example, usefuldetectable moieties include ³²P, fluorescent dyes, electron-densereagents, enzymes (e.g., as commonly used in an ELISA), biotin,digoxigenin, paramagnetic molecules, paramagnetic nanoparticles,ultrasmall superparamagnetic iron oxide (“USPIO”) nanoparticles, USPIOnanoparticle aggregates, superparamagnetic iron oxide (“SPIO”)nanoparticles, SPIO nanoparticle aggregates, monochrystalline SPIO,monochrystalline SPIO aggregates, monochrystalline iron oxidenanoparticles, monochrystalline iron oxide, other nanoparticle contrastagents, liposomes or other delivery vehicles containing Gadoliniumchelate (“Gd-chelate”) molecules, Gadolinium, radioisotopes,radionuclides (e.g. carbon-11, nitrogen-13, oxygen-15, fluorine-18,rubidium-82), fluorodeoxyglucose (e.g. fluorine-18 labeled), any gammaray emitting radionuclides, positron-emitting radionuclide, radiolabeledglucose, radiolabeled water, radiolabeled ammonia, biocolloids,microbubbles (e.g. including microbubble shells including albumin,galactose, lipid, and/or polymers; microbubble gas core including air,heavy gas(es), perfluorcarbon, nitrogen, octafluoropropane, perflexanelipid microsphere, perflutren, etc.), iodinated contrast agents (e.g.iohexol, iodixanol, ioversol, iopamidol, ioxilan, iopromide,diatrizoate, metrizoate, ioxaglate), barium sulfate, thorium dioxide,gold, gold nanoparticles, gold nanoparticle aggregates, fluorophores,two-photon fluorophores, or haptens and proteins or other entities whichcan be made detectable, e.g., by incorporating a radiolabel into apeptide or antibody specifically reactive with a target peptide.Detectable moieties also include any of the above compositionsencapsulated in nanoparticles, particles, aggregates, coated withadditional compositions, derivatized for binding to a targeting agent(e.g. compound described herein). Any method known in the art forconjugating an oligonucleotide or protein to the label may be employed,e.g., using methods described in Hermanson, Bioconiugate Techniques1996, Academic Press, Inc., San Diego.

MRI can be used to non-invasively acquire tissue images with highresolution. Paramagnetic agents or USPIO nanoparticles or aggregatesthereof enhance signal attenuation on T₂-weighted magnetic resonanceimages, and conjugation of such nanoparticles to binding ligands permitsthe detection of specific molecules at the cellular level. For example,MRI with nanoparticle detection agents can detect small foci of cancer.See e.g., Y. W. Jun et al., 2005, J. Am. Chern. Soc. 127:5732-5733; Y.M. Huh et al., 2005, J. Am. Chern. Soc. 127:12387-12391.Contrast-enhanced MRI is well-suited for the dynamic non-invasiveimaging of macromolecules or of molecular events, but it requiresligands that specifically bind to the molecule of interest. J. W. Bulteet al., 2004, NMR Biomed. 17:484-499. Fluorescent dyes and fluorophores(e.g. fluorescein, fluorescein isothiocyanate, and fluoresceinderivatives) can be used to non-invasively acquire tissue images withhigh resolution, with for example spectrophotometry, two-photonfluorescence, two-photon laser microscopy, or fluorescence microscopy(e.g. of tissue biopsies). MRI can be used to non-invasively acquiretissue images with high resolution, with for example paramagneticmolecules, paramagnetic nanoparticles, ultrasmall superparamagnetic ironoxide (“USPIO”) nanoparticles, USPIO nanoparticle aggregates,superparamagnetic iron oxide (“SPIO”) nanoparticles, SPIO nanoparticleaggregates, monochrystalline iron oxide nanoparticles, monochrystallineiron oxide, other nanoparticle contrast agents. MRI can be used tonon-invasively acquire tissue images with high resolution, with forexample Gadolinium, including liposomes or other delivery vehiclescontaining Gadolinium chelate (“Gd-chelate”) molecules. Positronemission tomography (PET), PET/computed tomography (CT), single photonemission computed tomography (SPECT), and SPECT/CT can be used tonon-invasively acquire tissue images with high resolution, with forexample radionuclides (e.g. carbon-11, nitrogen-13, oxygen-15,fluorine-18, rubidium-82), fluorodeoxyglucose (e.g. fluorine-18labeled), any gamma ray emitting radionuclides, positron-emittingradionuclide, radiolabeled glucose, radiolabeled water, radiolabeledammonia. Ultrasound (ultrasonography) and contrast enhanced ultrasound(contrast enhanced ultrasonography) can be used to non-invasivelyacquire tissue images with high resolution, with for example biocolloidsor microbubbles (e.g. including microbubble shells including albumin,galactose, lipid, and/or polymers; microbubble gas core including air,heavy gas(es), perfluorcarbon, nitrogen, octafluoropropane, perflexanelipid microsphere, perflutren, etc.). X-ray imaging (radiography) or CTcan be used to non-invasively acquire tissue images with highresolution, with for example iodinated contrast agents (e.g. iohexol,iodixanol, ioversol, iopamidol, ioxilan, iopromide, diatrizoate,metrizoate, ioxaglate), barium sulfate, thorium dioxide, gold, goldnanoparticles, or gold nanoparticle aggregates. These detection methodsand instruments and detectable moieties capable of being measured ordetected by the corresponding method are non-limiting examples.

As used herein, the term “conjugated” when referring to two moietiesmeans the two moieties are bonded, wherein the bond or bonds connectingthe two moieties may be covalent or non-covalent. In embodiments, thetwo moieties are covalently bonded to each other (e.g. directly orthrough a covalently bonded intermediary). In embodiments, the twomoieties are non-covalently bonded (e.g. through ionic bond(s), van derwaal's bond(s)/interactions, hydrogen bond(s), polar bond(s), orcombinations or mixtures thereof).

As used herein, the term “about” means a range of values including thespecified value, which a person of ordinary skill in the art wouldconsider reasonably similar to the specified value. In embodiments,about means within a standard deviation using measurements generallyacceptable in the art. In embodiments, about means a range extending to+/−10% of the specified value. In embodiments, about means the specifiedvalue.

II. Compositions

The nucleic acid polymer described herein can be generated viagraft-through polymerization, i.e., where the monomer is a“macromonomer” and is polymerized directly such that each positioncontains the side chain (e.g., nucleic acid) associated with themonomer. Such a polymer is entirely unique from analogues generated viapost-polymerization conjugation reactions since not every position ofthe analogue is modified. Polymers generated via graft-throughpolymerization and polymers generated via post-polymerizationmodification are fundamentally different structures. For instances,nucleic acid polymers of the present invention possess a dense array ofnucleic acids.

Graft-through polymerization can be performed by any method amenable tothe functional groups present. For instance, any polymerization strategycan be used to generate a dense, brush polymer accessible only viagraft-through polymerization of a monomer of nucleic acid.

In an aspect is provided a graft polymer including a linear backbonecovalently bound to a plurality of oligonucleotide branches. The graftpolymer is assembled by graft-through polymerization of a plurality ofoligonucleotide monomers including a polymerizable monomer covalentlybound to an oligonucleotide. The oligonucleotide thereby forming each ofthe plurality of oligonucleotide branches.

In embodiments, the graft-through polymerization employs ring-openingmetathesis polymerization (ROMP). In embodiments, the graft-throughpolymerization includes ring-opening metathesis polymerization (ROMP).In embodiments, the graft-through polymerization employs radicalpolymerization, controlled radical polymerization, reversibleaddition-fragmentation chain transfer (RAFT) polymerization, atomtransfer radical polymerization (ATRP), ring-opening metathesispolymerization (ROMP), anionic polymerization, cationic polymerization,free radical living polymerization, acyclic diene metathesispolymerization, radiation-induced polymerization, ring-opening olefinmetathesis polymerization, polycondensation reactions, oriniferter-induced polymerization.

In embodiments, the oligonucleotide (e.g. R³ or R⁴) includes at least 2different nucleobases. In embodiments, the oligonucleotide (e.g. R³ orR⁴) includes at least 3 different nucleobases. In embodiments, theoligonucleotide (e.g. R³ or R⁴) includes at least 4 differentnucleobases. In embodiments, the oligonucleotide (e.g. R³ or R⁴)includes at least 5 different nucleobases. In embodiments, theoligonucleotide (e.g. R³ or R⁴) includes at least 6 differentnucleobases. In embodiments, the oligonucleotide (e.g. R³ or R⁴)includes at least 7 different nucleobases. In embodiments, theoligonucleotide (e.g. R³ or R⁴) includes at least 3 nucleobases and atleast 2 different nucleobases. In embodiments, the oligonucleotide (e.g.R³ or R⁴) includes at least 5 nucleobases and at least 3 differentnucleobases. In embodiments, the oligonucleotide (e.g. R³ or R⁴)includes at least 10 nucleobases and at least 4 different nucleobases.In embodiments, the graft polymer includes at least 3 oligonucleotide(e.g. R³ or R⁴) branches. In embodiments, the graft polymer includes atleast 5 oligonucleotide (e.g. R³ or R⁴) branches. In embodiments, thegraft polymer includes at least 10 oligonucleotide (e.g. R³ or R⁴)branches. In embodiments, the oligonucleotide (e.g. R³ or R⁴) is between2 and 1000 bases long. In embodiments, the oligonucleotide is between 2and 900 bases long. In embodiments, the oligonucleotide is between 2 and800 bases long. In embodiments, the oligonucleotide is between 2 and 700bases long. In embodiments, the oligonucleotide is between 2 and 600bases long. In embodiments, the oligonucleotide is between 2 and 500bases long. In embodiments, the oligonucleotide is between 2 and 400bases long. In embodiments, the oligonucleotide is between 2 and 300bases long. In embodiments, the oligonucleotide is between 2 and 200bases long. In embodiments, the oligonucleotide is between 2 and 100bases long. In embodiments, the oligonucleotide is between 2 and 50bases long. In embodiments, the oligonucleotide is between 2 and 49bases long. In embodiments, the oligonucleotide is between 2 and 48bases long. In embodiments, the oligonucleotide is between 2 and 47bases long. In embodiments, the oligonucleotide is between 2 and 46bases long. In embodiments, the oligonucleotide is between 2 and 45bases long. In embodiments, the oligonucleotide is between 2 and 44bases long. In embodiments, the oligonucleotide is between 2 and 43bases long. In embodiments, the oligonucleotide is between 2 and 42bases long. In embodiments, the oligonucleotide is between 2 and 41bases long. In embodiments, the oligonucleotide is between 2 and 40bases long. In embodiments, the oligonucleotide is between 2 and 39bases long. In embodiments, the oligonucleotide is between 2 and 38bases long. In embodiments, the oligonucleotide is between 2 and 37bases long. In embodiments, the oligonucleotide is between 2 and 36bases long. In embodiments, the oligonucleotide is between 2 and 35bases long. In embodiments, the oligonucleotide is between 2 and 34bases long. In embodiments, the oligonucleotide is between 2 and 33bases long. In embodiments, the oligonucleotide is between 2 and 32bases long. In embodiments, the oligonucleotide is between 2 and 31bases long. In embodiments, the oligonucleotide is between 2 and 30bases long. In embodiments, the oligonucleotide is between 2 and 29bases long. In embodiments, the oligonucleotide is between 2 and 28bases long. In embodiments, the oligonucleotide is between 2 and 27bases long. In embodiments, the oligonucleotide is between 2 and 26bases long. In embodiments, the oligonucleotide is between 2 and 25bases long. In embodiments, the oligonucleotide is between 2 and 24bases long. In embodiments, the oligonucleotide is between 2 and 23bases long. In embodiments, the oligonucleotide is between 2 and 22bases long. In embodiments, the oligonucleotide is between 2 and 21bases long. In embodiments, the oligonucleotide is between 2 and 20bases long. In embodiments, the oligonucleotide is between 2 and 19bases long. In embodiments, the oligonucleotide is between 2 and 18bases long. In embodiments, the oligonucleotide is between 2 and 17bases long. In embodiments, the oligonucleotide is between 2 and 16bases long. In embodiments, the oligonucleotide is between 2 and 15bases long. In embodiments, the oligonucleotide is between 2 and 14bases long. In embodiments, the oligonucleotide is between 2 and 13bases long. In embodiments, the oligonucleotide is between 2 and 12bases long. In embodiments, the oligonucleotide is between 2 and 11bases long. In embodiments, the oligonucleotide is between 2 and 10bases long. In embodiments, the oligonucleotide is between 2 and 9 baseslong. In embodiments, the oligonucleotide is between 2 and 8 bases long.In embodiments, the oligonucleotide is between 2 and 7 bases long. Inembodiments, the oligonucleotide is between 2 and 6 bases long. Inembodiments, the oligonucleotide is between 2 and 5 bases long. Inembodiments, the oligonucleotide is between 2 and 4 bases long. Inembodiments, the oligonucleotide is between 2 and 3 bases long. Inembodiments, the oligonucleotide is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65,66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226,227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240,241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254,255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268,269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282,283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296,297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310,311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324,325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338,339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352,353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366,367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380,381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394,395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408,409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422,423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436,437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448, 449, 450,451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463, 464,465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478,479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492,493, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504, 505, 506,507, 508, 509, 510, 511, 512, 513, 514, 515, 516, 517, 518, 519, 520,521, 522, 523, 524, 525, 526, 527, 528, 529, 530, 531, 532, 533, 534,535, 536, 537, 538, 539, 540, 541, 542, 543, 544, 545, 546, 547, 548,549, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560, 561, 562,563, 564, 565, 566, 567, 568, 569, 570, 571, 572, 573, 574, 575, 576,577, 578, 579, 580, 581, 582, 583, 584, 585, 586, 587, 588, 589, 590,591, 592, 593, 594, 595, 596, 597, 598, 599, 600, 601, 602, 603, 604,605, 606, 607, 608, 609, 610, 611, 612, 613, 614, 615, 616, 617, 618,619, 620, 621, 622, 623, 624, 625, 626, 627, 628, 629, 630, 631, 632,633, 634, 635, 636, 637, 638, 639, 640, 641, 642, 643, 644, 645, 646,647, 648, 649, 650, 651, 652, 653, 654, 655, 656, 657, 658, 659, 660,661, 662, 663, 664, 665, 666, 667, 668, 669, 670, 671, 672, 673, 674,675, 676, 677, 678, 679, 680, 681, 682, 683, 684, 685, 686, 687, 688,689, 690, 691, 692, 693, 694, 695, 696, 697, 698, 699, 700, 701, 702,703, 704, 705, 706, 707, 708, 709, 710, 711, 712, 713, 714, 715, 716,717, 718, 719, 720, 721, 722, 723, 724, 725, 726, 727, 728, 729, 730,731, 732, 733, 734, 735, 736, 737, 738, 739, 740, 741, 742, 743, 744,745, 746, 747, 748, 749, 750, 751, 752, 753, 754, 755, 756, 757, 758,759, 760, 761, 762, 763, 764, 765, 766, 767, 768, 769, 770, 771, 772,773, 774, 775, 776, 777, 778, 779, 780, 781, 782, 783, 784, 785, 786,787, 788, 789, 790, 791, 792, 793, 794, 795, 796, 797, 798, 799, 800,801, 802, 803, 804, 805, 806, 807, 808, 809, 810, 811, 812, 813, 814,815, 816, 817, 818, 819, 820, 821, 822, 823, 824, 825, 826, 827, 828,829, 830, 831, 832, 833, 834, 835, 836, 837, 838, 839, 840, 841, 842,843, 844, 845, 846, 847, 848, 849, 850, 851, 852, 853, 854, 855, 856,857, 858, 859, 860, 861, 862, 863, 864, 865, 866, 867, 868, 869, 870,871, 872, 873, 874, 875, 876, 877, 878, 879, 880, 881, 882, 883, 884,885, 886, 887, 888, 889, 890, 891, 892, 893, 894, 895, 896, 897, 898,899, 900, 901, 902, 903, 904, 905, 906, 907, 908, 909, 910, 911, 912,913, 914, 915, 916, 917, 918, 919, 920, 921, 922, 923, 924, 925, 926,927, 928, 929, 930, 931, 932, 933, 934, 935, 936, 937, 938, 939, 940,941, 942, 943, 944, 945, 946, 947, 948, 949, 950, 951, 952, 953, 954,955, 956, 957, 958, 959, 960, 961, 962, 963, 964, 965, 966, 967, 968,969, 970, 971, 972, 973, 974, 975, 976, 977, 978, 979, 980, 981, 982,983, 984, 985, 986, 987, 988, 989, 990, 991, 992, 993, 994, 995, 996,997, 998, 999, or 1000 bases long.

In embodiments, the oligonucleotide (e.g. R³ or R⁴) includes nucleotideswith alternate backbones from the naturally occurring phophosphodiesterbond in DNA and RNA (e.g., phosphodiester derivatives). In embodiments,the oligonucleotide (e.g. R³ or R⁴) includes phosphoramidate,phosphorodiamidate, phosphorothioate (also known as phosphothioate),phosphorodithioate, phosphonocarboxylic acids, phosphonocarboxylates,phosphonoacetic acid, phosphonoformic acid, methyl phosphonate, boronphosphonate, peptide nucleic acid linkages, O-methylphosphoroamiditelinkages, positive backbone linkages; non-ionic backbone linkages,modified sugars, and/or non-ribose backbones (e.g. phosphorodiamidatemorpholino oligos or locked nucleic acids (LNA)). In embodiments, theoligonucleotide (e.g. R³ or R⁴) includes phosphodiester bonds such asthe naturally occurring backbone linkages in DNA or RNA.

In embodiments, the oligonucleotide includes phosphoramidate linkages.In embodiments, the oligonucleotide includes phosphorodiamidatelinkages. In embodiments, the oligonucleotide includes phosphorothioate(also known as phosphothioate) linkages. In embodiments, theoligonucleotide includes phosphorodithioate linkages. In embodiments,the oligonucleotide includes phosphonocarboxylic acid linkages. Inembodiments, the oligonucleotide includes phosphonocarboxylate linkages.In embodiments, the oligonucleotide includes phosphonoacetic acidlinkages. In embodiments, the oligonucleotide includes phosphonoformicacid linkages. In embodiments, the oligonucleotide includes methylphosphonate linkages. In embodiments, the oligonucleotide includes boronphosphonate linkages. In embodiments, the oligonucleotide includespeptide nucleic acid linkages. In embodiments, the oligonucleotideincludes O-methylphosphoroamidite linkages. In embodiments, theoligonucleotide includes positive backbone linkages. In embodiments, theoligonucleotide includes non-ionic backbone linkages. In embodiments,the oligonucleotide includes modified sugar linkages. In embodiments,the oligonucleotide includes non-ribose backbone linkages (e.g.phosphorodiamidate morpholino oligos or locked nucleic acids (LNA)).

In embodiments, the oligonucleotide (e.g. R³ or R⁴) includes aphosphoramidate linkage. In embodiments, the oligonucleotide includes aphosphorodiamidate linkage. In embodiments, the oligonucleotide includesa phosphorothioate (also known as phosphothioate) linkage. Inembodiments, the oligonucleotide includes a phosphorodithioate linkage.In embodiments, the oligonucleotide includes a phosphonocarboxylic acidlinkage. In embodiments, the oligonucleotide includes aphosphonocarboxylate linkage. In embodiments, the oligonucleotideincludes a phosphonoacetic acid linkage. In embodiments, theoligonucleotide includes a phosphonoformic acid linkage. In embodiments,the oligonucleotide includes a methyl phosphonate linkage. Inembodiments, the oligonucleotide includes a boron phosphonate linkage.In embodiments, the oligonucleotide includes a peptide nucleic acidlinkage. In embodiments, the oligonucleotide includes anO-methylphosphoroamidite linkage. In embodiments, the oligonucleotideincludes a positive backbone linkage. In embodiments, theoligonucleotide includes a non-ionic backbone linkage. In embodiments,the oligonucleotide includes a modified sugar linkage. In embodiments,the oligonucleotide includes a non-ribose backbone linkage (e.g.phosphorodiamidate morpholino oligos or locked nucleic acids (LNA)).

In embodiments, the graft polymer has the formula: R¹-[M(O)]_(n)—R². Thesymbol n is an integer from 2 to 1000. M is the polymerized product ofthe polymerizable monomer. O is the oligonucleotide. R¹ and R² areterminal polymer moieties.

In embodiments, n is an integer from 2 to 900. In embodiments, n is aninteger from 2 to 800. In embodiments, n is an integer from 2 to 700. Inembodiments, n is an integer from 2 to 600. In embodiments, n is aninteger from 2 to 500. In embodiments, n is an integer from 2 to 400. Inembodiments, n is an integer from 2 to 300. In embodiments, n is aninteger from 2 to 200. In embodiments, n is an integer from 2 to 100. Inembodiments, n is an integer from 2 to 50. In embodiments, n is aninteger from 2 to 49. In embodiments, n is an integer from 2 to 48. Inembodiments, n is an integer from 2 to 47. In embodiments, n is aninteger from 2 to 46. In embodiments, n is an integer from 2 to 45. Inembodiments, n is an integer from 2 to 44. In embodiments, n is aninteger from 2 to 43. In embodiments, n is an integer from 2 to 42. Inembodiments, n is an integer from 2 to 41. In embodiments, n is aninteger from 2 to 40. In embodiments, n is an integer from 2 to 39. Inembodiments, n is an integer from 2 to 38. In embodiments, n is aninteger from 2 to 37. In embodiments, n is an integer from 2 to 36. Inembodiments, n is an integer from 2 to 35. In embodiments, n is aninteger from 2 to 34. In embodiments, n is an integer from 2 to 33. Inembodiments, n is an integer from 2 to 32. In embodiments, n is aninteger from 2 to 31. In embodiments, n is an integer from 2 to 30. Inembodiments, n is an integer from 2 to 29. In embodiments, n is aninteger from 2 to 28. In embodiments, n is an integer from 2 to 27. Inembodiments, n is an integer from 2 to 26. In embodiments, n is aninteger from 2 to 25. In embodiments, n is an integer from 2 to 24. Inembodiments, n is an integer from 2 to 23. In embodiments, n is aninteger from 2 to 22. In embodiments, n is an integer from 2 to 21. Inembodiments, n is an integer from 2 to 20. In embodiments, n is aninteger from 2 to 19. In embodiments, n is an integer from 2 to 18. Inembodiments, n is an integer from 2 to 17. In embodiments, n is aninteger from 2 to 16. In embodiments, n is an integer from 2 to 15. Inembodiments, n is an integer from 2 to 14. In embodiments, n is aninteger from 2 to 13. In embodiments, n is an integer from 2 to 12. Inembodiments, n is an integer from 2 to 11. In embodiments, n is aninteger from 2 to 10. In embodiments, n is an integer from 2 to 9. Inembodiments, n is an integer from 2 to 8. In embodiments, n is aninteger from 2 to 7. In embodiments, n is an integer from 2 to 6. Inembodiments, n is an integer from 2 to 5. In embodiments, n is aninteger from 2 to 4. In embodiments, n is an integer from 2 to 3. Inembodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34,35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52,53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88,89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104,105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118,119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132,133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146,147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160,161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174,175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188,189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202,203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216,217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230,231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244,245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258,259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272,273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286,287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300,301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314,315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328,329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342,343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356,357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370,371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384,385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398,399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412,413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426,427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440,441, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454,455, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, 468,469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482,483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496,497, 498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510,511, 512, 513, 514, 515, 516, 517, 518, 519, 520, 521, 522, 523, 524,525, 526, 527, 528, 529, 530, 531, 532, 533, 534, 535, 536, 537, 538,539, 540, 541, 542, 543, 544, 545, 546, 547, 548, 549, 550, 551, 552,553, 554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564, 565, 566,567, 568, 569, 570, 571, 572, 573, 574, 575, 576, 577, 578, 579, 580,581, 582, 583, 584, 585, 586, 587, 588, 589, 590, 591, 592, 593, 594,595, 596, 597, 598, 599, 600, 601, 602, 603, 604, 605, 606, 607, 608,609, 610, 611, 612, 613, 614, 615, 616, 617, 618, 619, 620, 621, 622,623, 624, 625, 626, 627, 628, 629, 630, 631, 632, 633, 634, 635, 636,637, 638, 639, 640, 641, 642, 643, 644, 645, 646, 647, 648, 649, 650,651, 652, 653, 654, 655, 656, 657, 658, 659, 660, 661, 662, 663, 664,665, 666, 667, 668, 669, 670, 671, 672, 673, 674, 675, 676, 677, 678,679, 680, 681, 682, 683, 684, 685, 686, 687, 688, 689, 690, 691, 692,693, 694, 695, 696, 697, 698, 699, 700, 701, 702, 703, 704, 705, 706,707, 708, 709, 710, 711, 712, 713, 714, 715, 716, 717, 718, 719, 720,721, 722, 723, 724, 725, 726, 727, 728, 729, 730, 731, 732, 733, 734,735, 736, 737, 738, 739, 740, 741, 742, 743, 744, 745, 746, 747, 748,749, 750, 751, 752, 753, 754, 755, 756, 757, 758, 759, 760, 761, 762,763, 764, 765, 766, 767, 768, 769, 770, 771, 772, 773, 774, 775, 776,777, 778, 779, 780, 781, 782, 783, 784, 785, 786, 787, 788, 789, 790,791, 792, 793, 794, 795, 796, 797, 798, 799, 800, 801, 802, 803, 804,805, 806, 807, 808, 809, 810, 811, 812, 813, 814, 815, 816, 817, 818,819, 820, 821, 822, 823, 824, 825, 826, 827, 828, 829, 830, 831, 832,833, 834, 835, 836, 837, 838, 839, 840, 841, 842, 843, 844, 845, 846,847, 848, 849, 850, 851, 852, 853, 854, 855, 856, 857, 858, 859, 860,861, 862, 863, 864, 865, 866, 867, 868, 869, 870, 871, 872, 873, 874,875, 876, 877, 878, 879, 880, 881, 882, 883, 884, 885, 886, 887, 888,889, 890, 891, 892, 893, 894, 895, 896, 897, 898, 899, 900, 901, 902,903, 904, 905, 906, 907, 908, 909, 910, 911, 912, 913, 914, 915, 916,917, 918, 919, 920, 921, 922, 923, 924, 925, 926, 927, 928, 929, 930,931, 932, 933, 934, 935, 936, 937, 938, 939, 940, 941, 942, 943, 944,945, 946, 947, 948, 949, 950, 951, 952, 953, 954, 955, 956, 957, 958,959, 960, 961, 962, 963, 964, 965, 966, 967, 968, 969, 970, 971, 972,973, 974, 975, 976, 977, 978, 979, 980, 981, 982, 983, 984, 985, 986,987, 988, 989, 990, 991, 992, 993, 994, 995, 996, 997, 998, 999, or1000.

In embodiments, R¹ includes a solid support. In embodiments, R¹ includesa nanoparticle. In embodiments, R¹ includes a substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl or substituted orunsubstituted heteroaryl. In embodiments, R¹ includes a functionalmoiety. In embodiments, R¹ includes a detectable moiety. In embodiments,R¹ includes a ³²P, fluorescent dye, electron-dense reagent, enzyme(e.g., as commonly used in an ELISA), biotin, digoxigenin, paramagneticmolecule, paramagnetic nanoparticle, ultrasmall superparamagnetic ironoxide (“USPIO”) nanoparticles, USPIO nanoparticle aggregate,superparamagnetic iron oxide (“SPIO”) nanoparticle, SPIO nanoparticleaggregate, monochrystalline SPIO, monochrystalline SPIO aggregate,monochrystalline iron oxide nanoparticle, monochrystalline iron oxide,other nanoparticle contrast agent, liposome or other delivery vehiclecontaining Gadolinium chelate (“Gd-chelate”) molecule, Gadolinium,radioisotope, radionuclide (e.g. carbon-11, nitrogen-13, oxygen-15,fluorine-18, rubidium-82), fluorodeoxyglucose (e.g. fluorine-18labeled), any gamma ray emitting radionuclids, positron-emittingradionuclide, radiolabeled glucose, radiolabeled water, radiolabeledammonia, biocolloids, microbubble (e.g. including microbubble shellincluding albumin, galactose, lipid, and/or polymers; microbubble gascore including air, heavy gas(es), perfluorcarbon, nitrogen,octafluoropropane, perflexane lipid microsphere, perflutren, etc.),iodinated contrast agent (e.g. iohexol, iodixanol, ioversol, iopamidol,ioxilan, iopromide, diatrizoate, metrizoate, ioxaglate), barium sulfate,thorium dioxide, gold, gold nanoparticle, gold nanoparticle aggregate,fluorophore, two-photon fluorophore, or a hapten. In embodiments, R¹includes a polymerization product of an ethyl vinyl ether. Inembodiments, R¹ is the polymerization product of an alkene containingsubstituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl orsubstituted or unsubstituted heteroaryl. In embodiments, R¹ is thepolymerization product of an alkene bonded to a substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl or substituted orunsubstituted heteroaryl. In embodiments, R¹ is the polymerizationproduct of an alkene containing compound (e.g., also including afunction group, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, substituted or unsubstituted heteroaryl, ordetectable moiety). In embodiments, R¹ is selected from:

In embodiments, R² includes a solid support. In embodiments, R² includesa nanoparticle. In embodiments, R² includes a substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl or substituted orunsubstituted heteroaryl. In embodiments, R² includes a functionalmoiety. In embodiments, R² includes a detectable moiety. In embodiments,R² includes a ³²P, fluorescent dye, electron-dense reagent, enzyme(e.g., as commonly used in an ELISA), biotin, digoxigenin, paramagneticmolecule, paramagnetic nanoparticle, ultrasmall superparamagnetic ironoxide (“USPIO”) nanoparticles, USPIO nanoparticle aggregate,superparamagnetic iron oxide (“SPIO”) nanoparticle, SPIO nanoparticleaggregate, monochrystalline SPIO, monochrystalline SPIO aggregate,monochrystalline iron oxide nanoparticle, monochrystalline iron oxide,other nanoparticle contrast agent, liposome or other delivery vehiclecontaining Gadolinium chelate (“Gd-chelate”) molecule, Gadolinium,radioisotope, radionuclide (e.g. carbon-11, nitrogen-13, oxygen-15,fluorine-18, rubidium-82), fluorodeoxyglucose (e.g. fluorine-18labeled), any gamma ray emitting radionuclids, positron-emittingradionuclide, radiolabeled glucose, radiolabeled water, radiolabeledammonia, biocolloids, microbubble (e.g. including microbubble shellincluding albumin, galactose, lipid, and/or polymers; microbubble gascore including air, heavy gas(es), perfluorcarbon, nitrogen,octafluoropropane, perflexane lipid microsphere, perflutren, etc.),iodinated contrast agent (e.g. iohexol, iodixanol, ioversol, iopamidol,ioxilan, iopromide, diatrizoate, metrizoate, ioxaglate), barium sulfate,thorium dioxide, gold, gold nanoparticle, gold nanoparticle aggregate,fluorophore, two-photon fluorophore, or a hapten. In embodiments, R²includes a polymerization product of an ethyl vinyl ether. Inembodiments, R² is the polymerization product of an alkene containingsubstituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl orsubstituted or unsubstituted heteroaryl. In embodiments, R² is thepolymerization product of an alkene bonded to a substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl or substituted orunsubstituted heteroaryl. In embodiments, R² is the polymerizationproduct of an alkene containing compound (e.g., also including afunction group, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, substituted or unsubstituted heteroaryl, ordetectable moiety). In embodiments, R² is selected from:

In embodiments, the linear backbone is a polynorbornyl chain. Inembodiments, the linear backbone is a polynorbornyl derivative chain. Inembodiments, the linear backbone is a poly-substituted norbornyl chain.In embodiments, the linear backbone is a substituted polynorbornene. Inembodiments, the linear backbone is a polynorbornene. In embodiments,the linear backbone is a polynorbornene substituted witholigonucleotides at each norbornene monomer. In embodiments, the linearbackbone is a polynorbornene substituted with an oligonucleotide,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl and/orsubstituted or unsubstituted heteroaryl. In embodiments, the linkerbackbone is polymerized from monomers of:

wherein Ring A, n and R³ are as set forth herein. In embodiments, thelinker backbone is polymerized from monomers of:

In formula (IIA)-(IIF), Ring A, R³, R⁴, and L¹ are as set forth herein.In embodiments, the linker backbone is polymerized from monomers of:

In formula (IIIA)-(IIIF), Ring A, L¹ and R4 is as defined herein. Inembodiments, M(O) is

In embodiments, M(O) is

In embodiments, M(O) is

In embodiments, M(O) is

In embodiments, M(O) is

In embodiments, M(O) is

In embodiments, each oligonucleotide (e.g. R³ or R⁴) in the graftpolymer is optionally different. In embodiments, each oligonucleotide(e.g. R³ or R⁴) in the graft polymer is identical. In embodiments, thegraft polymer includes blocks of oligonucleotide (e.g. R³ or R⁴) whereinthe nucleotides in each block are identical and the oligonucleotide(e.g. R³ or R⁴) in different blocks are optionally different. Inembodiments, the graft polymer includes blocks of oligonucleotide (e.g.R³ or R⁴) wherein the nucleotides in each block are identical and theoligonucleotide (e.g. R³ or R⁴) in different blocks are different.

In an aspect is provided a block graft copolymer including a linearbackbone covalently bound to a plurality of oligonucleotide (e.g. R³ orR⁴) branches and a plurality of non-oligonucleotide side chains,wherein: the plurality of oligonucleotide branches form a first blockportion of the graft copolymer and the non-oligonucleotide side chainsform a second block portion of the graft copolymer; the graft copolymeris assembled by graft-through polymerization of a plurality ofoligonucleotide monomers and a plurality of non-oligonucleotidemonomers, wherein each of the plurality of oligonucleotide monomersincludes a polymerizable monomer covalently bound to an oligonucleotide,the oligonucleotide thereby forming each of the plurality ofoligonucleotide branches; and each of the plurality ofnon-oligonucleotide monomers includes the polymerizable monomercovalently bound to a non-oligonucleotide moiety, thenon-oligonucleotide moiety thereby forming each of the plurality ofnon-oligonucleotide side chains.

A “non-oligonucleotide monomer” is a polymerizable monomer that does notinclude an oligonucleotide. A non-oligonucleotide monomer may be apolymerizable monomer covalently bound to a substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, or substituted or unsubstitutedheteroaryl. In embodiments, a non-oligonucleotide monomer is ahydrophobic monomer. In embodiments, each non-oligonucleotide monomer inthe graft polymer is optionally different. In embodiments, eachnon-oligonucleotide monomer in the graft polymer is identical. Inembodiments, the graft polymer includes blocks of non-oligonucleotidemonomers wherein the non-oligonucleotide monomers in each block areidentical and the non-oligonucleotide monomers in different blocks areoptionally different. In embodiments, the graft polymer includes blocksof non-oligonucleotide monomers wherein the non-oligonucleotide monomersin each block are identical and the non-oligonucleotide monomers indifferent blocks are different. In embodiments, the non-oligonucleotidemonomer is selected from:

In embodiments, a non-oligonucleotide monomer is a hydrophobic monomer.

In an aspect is provided an amphiphilic block graft copolymer includinga linear backbone covalently bound to a plurality of oligonucleotide(e.g. R³ or R⁴) branches and a plurality of hydrophobic side chains,wherein: the plurality of oligonucleotide branches form a hydrophilicblock portion of the amphiphilic graft copolymer and the hydrophobicside chains form a hydrophobic block portion of the amphiphilic graftcopolymer; the graft copolymer is assembled by graft-throughpolymerization of a plurality of oligonucleotide monomers and aplurality of hydrophobic monomers, wherein each of the plurality ofoligonucleotide monomers includes a polymerizable monomer covalentlybound to an oligonucleotide, the oligonucleotide thereby forming each ofthe plurality of oligonucleotide branches; and each of the plurality ofhydrophobic monomers includes the polymerizable monomer covalently boundto a hydrophobic moiety, the hydrophobic moiety thereby forming each ofthe plurality of hydrophobic side chains.

The linear backbone, oligonucleotide (e.g. R³ or R⁴) branches,oligonucleotide monomers, graft-through polymerization, polymerizablemonomer, and oligonucleotide are as described herein, including inaspects (e.g., above), embodiments (e.g., above), examples, figures,tables, schemes, and claims.

In embodiments, the hydrophobic moiety is sufficiently hydrophobic andof sufficient size such that the amphiphilic block graft polymer iscapable of forming a micelle in an aqueous-based solvent. Inembodiments, the hydrophobic moiety is a substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, or substituted or unsubstitutedheteroaryl. In embodiments, the hydrophobic moiety is an unsubstitutedbenzyl. In embodiments, the hydrophobic moiety is a hydrophobic moietyas shown in FIG. 2.

In embodiments, the amphiphilic block graft copolymer has the formula:R¹-[M(O)]_(n)-[M(P)]_(m)—R² or R¹-[M(P)]_(m)-[M(O)]_(n)—R² wherein, n isan integer from 2 to 1000; m is an integer from 2 to 1000; M is thepolymerized product of the polymerizable monomer; O is theoligonucleotide; P is the hydrophobic moiety; and R¹ and R² are terminalpolymer moieties.

R¹, M, O, n, and R² are as described herein, including in aspects (e.g.,above), embodiments (e.g., above), examples, figures, tables, schemes,and claims. In embodiments, M(P) is

In embodiments, M(P) is

In embodiments, M(P) is

In embodiments, M(P) is

In embodiments, M(P) is

In embodiments, M(P) is

In embodiments, M(P) is

In embodiments, each hydrophobic moiety in the graft copolymer isoptionally different. In embodiments, each hydrophobic moiety in thegraft copolymer is identical. In embodiments, the graft copolymerincludes blocks of hydrophobic moieties wherein the hydrophobic moietiesin each block are identical and the hydrophobic moieties in differentblocks are optionally different. In embodiments, the graft copolymerincludes blocks of hydrophobic moieties wherein the hydrophobic moietiesin each block are identical and the hydrophobic moieties in differentblocks are different.

In embodiments, m is an integer from 2 to 900. In embodiments, m is aninteger from 2 to 800. In embodiments, m is an integer from 2 to 700. Inembodiments, m is an integer from 2 to 600. In embodiments, m is aninteger from 2 to 500. In embodiments, m is an integer from 2 to 400. Inembodiments, m is an integer from 2 to 300. In embodiments, m is aninteger from 2 to 200. In embodiments, m is an integer from 2 to 100. Inembodiments, m is an integer from 2 to 50. In embodiments, m is aninteger from 2 to 49. In embodiments, m is an integer from 2 to 48. Inembodiments, m is an integer from 2 to 47. In embodiments, m is aninteger from 2 to 46. In embodiments, m is an integer from 2 to 45. Inembodiments, m is an integer from 2 to 44. In embodiments, m is aninteger from 2 to 43. In embodiments, m is an integer from 2 to 42. Inembodiments, m is an integer from 2 to 41. In embodiments, m is aninteger from 2 to 40. In embodiments, m is an integer from 2 to 39. Inembodiments, m is an integer from 2 to 38. In embodiments, m is aninteger from 2 to 37. In embodiments, m is an integer from 2 to 36. Inembodiments, m is an integer from 2 to 35. In embodiments, m is aninteger from 2 to 34. In embodiments, m is an integer from 2 to 33. Inembodiments, m is an integer from 2 to 32. In embodiments, m is aninteger from 2 to 31. In embodiments, m is an integer from 2 to 30. Inembodiments, m is an integer from 2 to 29. In embodiments, m is aninteger from 2 to 28. In embodiments, m is an integer from 2 to 27. Inembodiments, m is an integer from 2 to 26. In embodiments, m is aninteger from 2 to 25. In embodiments, m is an integer from 2 to 24. Inembodiments, m is an integer from 2 to 23. In embodiments, m is aninteger from 2 to 22. In embodiments, m is an integer from 2 to 21. Inembodiments, m is an integer from 2 to 20. In embodiments, m is aninteger from 2 to 19. In embodiments, m is an integer from 2 to 18. Inembodiments, m is an integer from 2 to 17. In embodiments, m is aninteger from 2 to 16. In embodiments, m is an integer from 2 to 15. Inembodiments, m is an integer from 2 to 14. In embodiments, m is aninteger from 2 to 13. In embodiments, m is an integer from 2 to 12. Inembodiments, m is an integer from 2 to 11. In embodiments, m is aninteger from 2 to 10. In embodiments, m is an integer from 2 to 9. Inembodiments, m is an integer from 2 to 8. In embodiments, m is aninteger from 2 to 7. In embodiments, m is an integer from 2 to 6. Inembodiments, m is an integer from 2 to 5. In embodiments, m is aninteger from 2 to 4. In embodiments, m is an integer from 2 to 3. Inembodiments, m is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34,35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52,53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88,89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104,105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118,119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132,133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146,147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160,161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174,175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188,189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202,203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216,217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230,231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244,245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258,259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272,273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286,287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300,301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314,315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328,329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342,343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356,357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370,371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384,385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398,399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412,413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426,427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440,441, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454,455, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, 468,469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482,483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496,497, 498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510,511, 512, 513, 514, 515, 516, 517, 518, 519, 520, 521, 522, 523, 524,525, 526, 527, 528, 529, 530, 531, 532, 533, 534, 535, 536, 537, 538,539, 540, 541, 542, 543, 544, 545, 546, 547, 548, 549, 550, 551, 552,553, 554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564, 565, 566,567, 568, 569, 570, 571, 572, 573, 574, 575, 576, 577, 578, 579, 580,581, 582, 583, 584, 585, 586, 587, 588, 589, 590, 591, 592, 593, 594,595, 596, 597, 598, 599, 600, 601, 602, 603, 604, 605, 606, 607, 608,609, 610, 611, 612, 613, 614, 615, 616, 617, 618, 619, 620, 621, 622,623, 624, 625, 626, 627, 628, 629, 630, 631, 632, 633, 634, 635, 636,637, 638, 639, 640, 641, 642, 643, 644, 645, 646, 647, 648, 649, 650,651, 652, 653, 654, 655, 656, 657, 658, 659, 660, 661, 662, 663, 664,665, 666, 667, 668, 669, 670, 671, 672, 673, 674, 675, 676, 677, 678,679, 680, 681, 682, 683, 684, 685, 686, 687, 688, 689, 690, 691, 692,693, 694, 695, 696, 697, 698, 699, 700, 701, 702, 703, 704, 705, 706,707, 708, 709, 710, 711, 712, 713, 714, 715, 716, 717, 718, 719, 720,721, 722, 723, 724, 725, 726, 727, 728, 729, 730, 731, 732, 733, 734,735, 736, 737, 738, 739, 740, 741, 742, 743, 744, 745, 746, 747, 748,749, 750, 751, 752, 753, 754, 755, 756, 757, 758, 759, 760, 761, 762,763, 764, 765, 766, 767, 768, 769, 770, 771, 772, 773, 774, 775, 776,777, 778, 779, 780, 781, 782, 783, 784, 785, 786, 787, 788, 789, 790,791, 792, 793, 794, 795, 796, 797, 798, 799, 800, 801, 802, 803, 804,805, 806, 807, 808, 809, 810, 811, 812, 813, 814, 815, 816, 817, 818,819, 820, 821, 822, 823, 824, 825, 826, 827, 828, 829, 830, 831, 832,833, 834, 835, 836, 837, 838, 839, 840, 841, 842, 843, 844, 845, 846,847, 848, 849, 850, 851, 852, 853, 854, 855, 856, 857, 858, 859, 860,861, 862, 863, 864, 865, 866, 867, 868, 869, 870, 871, 872, 873, 874,875, 876, 877, 878, 879, 880, 881, 882, 883, 884, 885, 886, 887, 888,889, 890, 891, 892, 893, 894, 895, 896, 897, 898, 899, 900, 901, 902,903, 904, 905, 906, 907, 908, 909, 910, 911, 912, 913, 914, 915, 916,917, 918, 919, 920, 921, 922, 923, 924, 925, 926, 927, 928, 929, 930,931, 932, 933, 934, 935, 936, 937, 938, 939, 940, 941, 942, 943, 944,945, 946, 947, 948, 949, 950, 951, 952, 953, 954, 955, 956, 957, 958,959, 960, 961, 962, 963, 964, 965, 966, 967, 968, 969, 970, 971, 972,973, 974, 975, 976, 977, 978, 979, 980, 981, 982, 983, 984, 985, 986,987, 988, 989, 990, 991, 992, 993, 994, 995, 996, 997, 998, 999, or1000.

In an aspect is provided a micelle including an amphiphilic block graftcopolymer described herein, including in an aspect, embodiment, example,figures, table, scheme, or claim.

In embodiments, the micelle has a diameter of between about 1 and about1000 nm. In embodiments, the micelle has a diameter of between about 5and about 100 nm. In embodiments, the micelle has a diameter of betweenabout 10 and about 50 nm. In embodiments, the micelle has a diameter ofbetween 1 and 1000 nm. In embodiments, the micelle has a diameter ofbetween 5 and 100 nm. In embodiments, the micelle has a diameter ofbetween 10 and 50 nm.

In embodiments, the micelle has a diameter of about 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61,62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79,80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224,225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238,239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252,253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266,267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280,281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294,295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308,309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322,323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336,337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350,351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364,365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378,379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392,393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406,407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420,421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434,435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448,449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462,463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476,477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490,491, 492, 493, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504,505, 506, 507, 508, 509, 510, 511, 512, 513, 514, 515, 516, 517, 518,519, 520, 521, 522, 523, 524, 525, 526, 527, 528, 529, 530, 531, 532,533, 534, 535, 536, 537, 538, 539, 540, 541, 542, 543, 544, 545, 546,547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560,561, 562, 563, 564, 565, 566, 567, 568, 569, 570, 571, 572, 573, 574,575, 576, 577, 578, 579, 580, 581, 582, 583, 584, 585, 586, 587, 588,589, 590, 591, 592, 593, 594, 595, 596, 597, 598, 599, 600, 601, 602,603, 604, 605, 606, 607, 608, 609, 610, 611, 612, 613, 614, 615, 616,617, 618, 619, 620, 621, 622, 623, 624, 625, 626, 627, 628, 629, 630,631, 632, 633, 634, 635, 636, 637, 638, 639, 640, 641, 642, 643, 644,645, 646, 647, 648, 649, 650, 651, 652, 653, 654, 655, 656, 657, 658,659, 660, 661, 662, 663, 664, 665, 666, 667, 668, 669, 670, 671, 672,673, 674, 675, 676, 677, 678, 679, 680, 681, 682, 683, 684, 685, 686,687, 688, 689, 690, 691, 692, 693, 694, 695, 696, 697, 698, 699, 700,701, 702, 703, 704, 705, 706, 707, 708, 709, 710, 711, 712, 713, 714,715, 716, 717, 718, 719, 720, 721, 722, 723, 724, 725, 726, 727, 728,729, 730, 731, 732, 733, 734, 735, 736, 737, 738, 739, 740, 741, 742,743, 744, 745, 746, 747, 748, 749, 750, 751, 752, 753, 754, 755, 756,757, 758, 759, 760, 761, 762, 763, 764, 765, 766, 767, 768, 769, 770,771, 772, 773, 774, 775, 776, 777, 778, 779, 780, 781, 782, 783, 784,785, 786, 787, 788, 789, 790, 791, 792, 793, 794, 795, 796, 797, 798,799, 800, 801, 802, 803, 804, 805, 806, 807, 808, 809, 810, 811, 812,813, 814, 815, 816, 817, 818, 819, 820, 821, 822, 823, 824, 825, 826,827, 828, 829, 830, 831, 832, 833, 834, 835, 836, 837, 838, 839, 840,841, 842, 843, 844, 845, 846, 847, 848, 849, 850, 851, 852, 853, 854,855, 856, 857, 858, 859, 860, 861, 862, 863, 864, 865, 866, 867, 868,869, 870, 871, 872, 873, 874, 875, 876, 877, 878, 879, 880, 881, 882,883, 884, 885, 886, 887, 888, 889, 890, 891, 892, 893, 894, 895, 896,897, 898, 899, 900, 901, 902, 903, 904, 905, 906, 907, 908, 909, 910,911, 912, 913, 914, 915, 916, 917, 918, 919, 920, 921, 922, 923, 924,925, 926, 927, 928, 929, 930, 931, 932, 933, 934, 935, 936, 937, 938,939, 940, 941, 942, 943, 944, 945, 946, 947, 948, 949, 950, 951, 952,953, 954, 955, 956, 957, 958, 959, 960, 961, 962, 963, 964, 965, 966,967, 968, 969, 970, 971, 972, 973, 974, 975, 976, 977, 978, 979, 980,981, 982, 983, 984, 985, 986, 987, 988, 989, 990, 991, 992, 993, 994,995, 996, 997, 998, 999, or 1000 nm. In embodiments, the micelle has adiameter of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53,54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105,106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119,120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133,134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147,148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161,162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175,176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189,190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203,204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217,218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231,232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245,246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259,260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273,274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287,288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301,302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315,316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329,330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343,344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357,358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371,372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385,386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399,400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413,414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427,428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441,442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454, 455,456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, 468, 469,470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483,484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497,498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, 511,512, 513, 514, 515, 516, 517, 518, 519, 520, 521, 522, 523, 524, 525,526, 527, 528, 529, 530, 531, 532, 533, 534, 535, 536, 537, 538, 539,540, 541, 542, 543, 544, 545, 546, 547, 548, 549, 550, 551, 552, 553,554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564, 565, 566, 567,568, 569, 570, 571, 572, 573, 574, 575, 576, 577, 578, 579, 580, 581,582, 583, 584, 585, 586, 587, 588, 589, 590, 591, 592, 593, 594, 595,596, 597, 598, 599, 600, 601, 602, 603, 604, 605, 606, 607, 608, 609,610, 611, 612, 613, 614, 615, 616, 617, 618, 619, 620, 621, 622, 623,624, 625, 626, 627, 628, 629, 630, 631, 632, 633, 634, 635, 636, 637,638, 639, 640, 641, 642, 643, 644, 645, 646, 647, 648, 649, 650, 651,652, 653, 654, 655, 656, 657, 658, 659, 660, 661, 662, 663, 664, 665,666, 667, 668, 669, 670, 671, 672, 673, 674, 675, 676, 677, 678, 679,680, 681, 682, 683, 684, 685, 686, 687, 688, 689, 690, 691, 692, 693,694, 695, 696, 697, 698, 699, 700, 701, 702, 703, 704, 705, 706, 707,708, 709, 710, 711, 712, 713, 714, 715, 716, 717, 718, 719, 720, 721,722, 723, 724, 725, 726, 727, 728, 729, 730, 731, 732, 733, 734, 735,736, 737, 738, 739, 740, 741, 742, 743, 744, 745, 746, 747, 748, 749,750, 751, 752, 753, 754, 755, 756, 757, 758, 759, 760, 761, 762, 763,764, 765, 766, 767, 768, 769, 770, 771, 772, 773, 774, 775, 776, 777,778, 779, 780, 781, 782, 783, 784, 785, 786, 787, 788, 789, 790, 791,792, 793, 794, 795, 796, 797, 798, 799, 800, 801, 802, 803, 804, 805,806, 807, 808, 809, 810, 811, 812, 813, 814, 815, 816, 817, 818, 819,820, 821, 822, 823, 824, 825, 826, 827, 828, 829, 830, 831, 832, 833,834, 835, 836, 837, 838, 839, 840, 841, 842, 843, 844, 845, 846, 847,848, 849, 850, 851, 852, 853, 854, 855, 856, 857, 858, 859, 860, 861,862, 863, 864, 865, 866, 867, 868, 869, 870, 871, 872, 873, 874, 875,876, 877, 878, 879, 880, 881, 882, 883, 884, 885, 886, 887, 888, 889,890, 891, 892, 893, 894, 895, 896, 897, 898, 899, 900, 901, 902, 903,904, 905, 906, 907, 908, 909, 910, 911, 912, 913, 914, 915, 916, 917,918, 919, 920, 921, 922, 923, 924, 925, 926, 927, 928, 929, 930, 931,932, 933, 934, 935, 936, 937, 938, 939, 940, 941, 942, 943, 944, 945,946, 947, 948, 949, 950, 951, 952, 953, 954, 955, 956, 957, 958, 959,960, 961, 962, 963, 964, 965, 966, 967, 968, 969, 970, 971, 972, 973,974, 975, 976, 977, 978, 979, 980, 981, 982, 983, 984, 985, 986, 987,988, 989, 990, 991, 992, 993, 994, 995, 996, 997, 998, 999, or 1000 nm.

In embodiments, the micelle diameter is a hydrodynamic diameter. Inembodiments, the diameter is an average diameter of a sample.

In an aspect is provided a nanoparticle including an amphiphilic blockgraft copolymer described herein, including in an aspect, embodiment,example, figure, table, scheme, or claim.

In embodiments, the nanoparticle is a spherical nanoparticle.

In embodiments, the nanoparticle has a diameter of between about 1 andabout 1000 nm. In embodiments, the nanoparticle has a diameter ofbetween about 5 and about 100 nm. In embodiments, the nanoparticle has adiameter of between about 10 and about 50 nm. In embodiments, thenanoparticle has a diameter of between 1 and 1000 nm. In embodiments,the nanoparticle has a diameter of between 5 and 100 nm. In embodiments,the nanoparticle has a diameter of between 10 and 50 nm.

In embodiments, the nanoparticle has a diameter of about 1, 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42,43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60,61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111,112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125,126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139,140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153,154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167,168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181,182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195,196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209,210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223,224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237,238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251,252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265,266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279,280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293,294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307,308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321,322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335,336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349,350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363,364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377,378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391,392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405,406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419,420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433,434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447,448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461,462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475,476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489,490, 491, 492, 493, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503,504, 505, 506, 507, 508, 509, 510, 511, 512, 513, 514, 515, 516, 517,518, 519, 520, 521, 522, 523, 524, 525, 526, 527, 528, 529, 530, 531,532, 533, 534, 535, 536, 537, 538, 539, 540, 541, 542, 543, 544, 545,546, 547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559,560, 561, 562, 563, 564, 565, 566, 567, 568, 569, 570, 571, 572, 573,574, 575, 576, 577, 578, 579, 580, 581, 582, 583, 584, 585, 586, 587,588, 589, 590, 591, 592, 593, 594, 595, 596, 597, 598, 599, 600, 601,602, 603, 604, 605, 606, 607, 608, 609, 610, 611, 612, 613, 614, 615,616, 617, 618, 619, 620, 621, 622, 623, 624, 625, 626, 627, 628, 629,630, 631, 632, 633, 634, 635, 636, 637, 638, 639, 640, 641, 642, 643,644, 645, 646, 647, 648, 649, 650, 651, 652, 653, 654, 655, 656, 657,658, 659, 660, 661, 662, 663, 664, 665, 666, 667, 668, 669, 670, 671,672, 673, 674, 675, 676, 677, 678, 679, 680, 681, 682, 683, 684, 685,686, 687, 688, 689, 690, 691, 692, 693, 694, 695, 696, 697, 698, 699,700, 701, 702, 703, 704, 705, 706, 707, 708, 709, 710, 711, 712, 713,714, 715, 716, 717, 718, 719, 720, 721, 722, 723, 724, 725, 726, 727,728, 729, 730, 731, 732, 733, 734, 735, 736, 737, 738, 739, 740, 741,742, 743, 744, 745, 746, 747, 748, 749, 750, 751, 752, 753, 754, 755,756, 757, 758, 759, 760, 761, 762, 763, 764, 765, 766, 767, 768, 769,770, 771, 772, 773, 774, 775, 776, 777, 778, 779, 780, 781, 782, 783,784, 785, 786, 787, 788, 789, 790, 791, 792, 793, 794, 795, 796, 797,798, 799, 800, 801, 802, 803, 804, 805, 806, 807, 808, 809, 810, 811,812, 813, 814, 815, 816, 817, 818, 819, 820, 821, 822, 823, 824, 825,826, 827, 828, 829, 830, 831, 832, 833, 834, 835, 836, 837, 838, 839,840, 841, 842, 843, 844, 845, 846, 847, 848, 849, 850, 851, 852, 853,854, 855, 856, 857, 858, 859, 860, 861, 862, 863, 864, 865, 866, 867,868, 869, 870, 871, 872, 873, 874, 875, 876, 877, 878, 879, 880, 881,882, 883, 884, 885, 886, 887, 888, 889, 890, 891, 892, 893, 894, 895,896, 897, 898, 899, 900, 901, 902, 903, 904, 905, 906, 907, 908, 909,910, 911, 912, 913, 914, 915, 916, 917, 918, 919, 920, 921, 922, 923,924, 925, 926, 927, 928, 929, 930, 931, 932, 933, 934, 935, 936, 937,938, 939, 940, 941, 942, 943, 944, 945, 946, 947, 948, 949, 950, 951,952, 953, 954, 955, 956, 957, 958, 959, 960, 961, 962, 963, 964, 965,966, 967, 968, 969, 970, 971, 972, 973, 974, 975, 976, 977, 978, 979,980, 981, 982, 983, 984, 985, 986, 987, 988, 989, 990, 991, 992, 993,994, 995, 996, 997, 998, 999, or 1000 nm.

In embodiments, the nanoparticle has a diameter of 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61,62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79,80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224,225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238,239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252,253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266,267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280,281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294,295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308,309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322,323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336,337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350,351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364,365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378,379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392,393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406,407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420,421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434,435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448,449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462,463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476,477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490,491, 492, 493, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504,505, 506, 507, 508, 509, 510, 511, 512, 513, 514, 515, 516, 517, 518,519, 520, 521, 522, 523, 524, 525, 526, 527, 528, 529, 530, 531, 532,533, 534, 535, 536, 537, 538, 539, 540, 541, 542, 543, 544, 545, 546,547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560,561, 562, 563, 564, 565, 566, 567, 568, 569, 570, 571, 572, 573, 574,575, 576, 577, 578, 579, 580, 581, 582, 583, 584, 585, 586, 587, 588,589, 590, 591, 592, 593, 594, 595, 596, 597, 598, 599, 600, 601, 602,603, 604, 605, 606, 607, 608, 609, 610, 611, 612, 613, 614, 615, 616,617, 618, 619, 620, 621, 622, 623, 624, 625, 626, 627, 628, 629, 630,631, 632, 633, 634, 635, 636, 637, 638, 639, 640, 641, 642, 643, 644,645, 646, 647, 648, 649, 650, 651, 652, 653, 654, 655, 656, 657, 658,659, 660, 661, 662, 663, 664, 665, 666, 667, 668, 669, 670, 671, 672,673, 674, 675, 676, 677, 678, 679, 680, 681, 682, 683, 684, 685, 686,687, 688, 689, 690, 691, 692, 693, 694, 695, 696, 697, 698, 699, 700,701, 702, 703, 704, 705, 706, 707, 708, 709, 710, 711, 712, 713, 714,715, 716, 717, 718, 719, 720, 721, 722, 723, 724, 725, 726, 727, 728,729, 730, 731, 732, 733, 734, 735, 736, 737, 738, 739, 740, 741, 742,743, 744, 745, 746, 747, 748, 749, 750, 751, 752, 753, 754, 755, 756,757, 758, 759, 760, 761, 762, 763, 764, 765, 766, 767, 768, 769, 770,771, 772, 773, 774, 775, 776, 777, 778, 779, 780, 781, 782, 783, 784,785, 786, 787, 788, 789, 790, 791, 792, 793, 794, 795, 796, 797, 798,799, 800, 801, 802, 803, 804, 805, 806, 807, 808, 809, 810, 811, 812,813, 814, 815, 816, 817, 818, 819, 820, 821, 822, 823, 824, 825, 826,827, 828, 829, 830, 831, 832, 833, 834, 835, 836, 837, 838, 839, 840,841, 842, 843, 844, 845, 846, 847, 848, 849, 850, 851, 852, 853, 854,855, 856, 857, 858, 859, 860, 861, 862, 863, 864, 865, 866, 867, 868,869, 870, 871, 872, 873, 874, 875, 876, 877, 878, 879, 880, 881, 882,883, 884, 885, 886, 887, 888, 889, 890, 891, 892, 893, 894, 895, 896,897, 898, 899, 900, 901, 902, 903, 904, 905, 906, 907, 908, 909, 910,911, 912, 913, 914, 915, 916, 917, 918, 919, 920, 921, 922, 923, 924,925, 926, 927, 928, 929, 930, 931, 932, 933, 934, 935, 936, 937, 938,939, 940, 941, 942, 943, 944, 945, 946, 947, 948, 949, 950, 951, 952,953, 954, 955, 956, 957, 958, 959, 960, 961, 962, 963, 964, 965, 966,967, 968, 969, 970, 971, 972, 973, 974, 975, 976, 977, 978, 979, 980,981, 982, 983, 984, 985, 986, 987, 988, 989, 990, 991, 992, 993, 994,995, 996, 997, 998, 999, or 1000 nm.

In embodiments, the nanoparticle diameter is a hydrodynamic diameter. Inembodiments, the diameter is an average diameter of a sample.

In embodiments, the graft polymer is capable of programmedself-assembly. In embodiments, the programmed self-assembly results in ananoparticle (e.g., as described herein).

In embodiments, the graft polymer is a probe for DNA recognition. Inembodiments, the graft polymer is a probe for nucleic acid recognition.In embodiments, the graft polymer is capable of intracellular genemanipulation (e.g., knockdown, inhibition, reduction). In embodiments,the graft polymer is a capable of RNA manipulation (e.g., knockdown,inhibition, reduction). In embodiments, the graft polymer is a capableof cellular internalization of nucleic acids. In embodiments, the graftpolymer is a capable of gene interference. In embodiments, the graftpolymer is a capable of theranostics. In embodiments, the graft polymeris a capable of cellular internalization.

Detailed descriptions of exemplary embodiments of the nucleic acidpolymer are provided in the examples section below and throughout thepresent application.

In embodiments, the graft polymer, micelle, amphiphilic block graftcopolymer, or nanoparticle is as described herein, including in anaspect, embodiment, example, figure, table, scheme, and claim.

III. Methods of Making Polymers

In an aspect is provided a method of making a graft polymer, the methodincluding: (i) reacting a plurality of oligonucleotide monomers with apolymerization catalyst or initiator, wherein each of the plurality ofoligonucleotide monomers includes a polymerizable monomer covalentlybound to an oligonucleotide; and (ii) terminating the reacting with achain terminator or transfer agent.

In embodiments, the graft polymer is a graft polymer described herein(including in an aspect, embodiment, example, figure, table, claim, orscheme). In embodiments, the graft polymer is a brush polymer.

In embodiments, the graft polymer includes a linear backbone covalentlybound to a plurality of oligonucleotide branches.

In embodiments, the polymerization catalyst or initiator is a ROMPcatalyst. In embodiments, the polymerization catalyst or initiator is aruthenium catalyst. In embodiments, the ROMP uses a Grubbs' catalyst. Inembodiments, the ROMP uses a Mo catalyst. In embodiments, thepolymerization catalyst or initiator is a [Mo(═CHBut)(Nar)(OR)2]. Inembodiments, the polymerization catalyst or initiator is a transitionmetal catalyst. In embodiments, the polymerization catalyst or initiatoris a transition metal carbine complex catalyst In embodiments, thepolymerization catalyst or initiator is aBenzylidene-bis(tricyclohexylphosphine)-dichlororuthenium. Inembodiments, the polymerization catalyst or initiator is a[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(phenylmethylene)(tricyclohexylphosphine)ruthenium.In embodiments, the polymerization catalyst or initiator is aDichloro(o-isopropoxyphenylmethylene)(tricyclohexylphosphine)ruthenium(II).In embodiments, the polymerization catalyst or initiator is a[1,3-Bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(o-isopropoxyphenylmethylene)ruthenium.In embodiments, the polymerization catalyst or initiator is a thirdgeneration Grubbs' catalyst. In embodiments, the polymerization catalystor initiator is a (IMesH₂)(C₅H₅N)₂(Cl)₂Ru═CHPh. In embodiments, thepolymerization catalyst or initiator is a radical generating compound.In embodiments, the polymerization catalyst or initiator is a freeradical compound.

In embodiments, the chain terminator or transfer agent includes a solidsupport. In embodiments, the chain terminator or transfer agent includesa nanoparticle. In embodiments, the chain terminator or transfer agentincludes a substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl or substituted or unsubstituted heteroaryl. Inembodiments, the chain terminator or transfer agent includes afunctional moiety. In embodiments, the chain terminator or transferagent includes a detectable moiety. In embodiments, the chain terminatoror transfer agent includes a ³²P, fluorescent dye, electron-densereagent, enzyme (e.g., as commonly used in an ELISA), biotin,digoxigenin, paramagnetic molecule, paramagnetic nanoparticle,ultrasmall superparamagnetic iron oxide (“USPIO”) nanoparticles, USPIOnanoparticle aggregate, superparamagnetic iron oxide (“SPIO”)nanoparticle, SPIO nanoparticle aggregate, monochrystalline SPIO,monochrystalline SPIO aggregate, monochrystalline iron oxidenanoparticle, monochrystalline iron oxide, other nanoparticle contrastagent, liposome or other delivery vehicle containing Gadolinium chelate(“Gd-chelate”) molecule, Gadolinium, radioisotope, radionuclide (e.g.carbon-11, nitrogen-13, oxygen-15, fluorine-18, rubidium-82),fluorodeoxyglucose (e.g. fluorine-18 labeled), any gamma ray emittingradionuclids, positron-emitting radionuclide, radiolabeled glucose,radiolabeled water, radiolabeled ammonia, biocolloids, microbubble (e.g.including microbubble shell including albumin, galactose, lipid, and/orpolymers; microbubble gas core including air, heavy gas(es),perfluorcarbon, nitrogen, octafluoropropane, perflexane lipidmicrosphere, perflutren, etc.), iodinated contrast agent (e.g. iohexol,iodixanol, ioversol, iopamidol, ioxilan, iopromide, diatrizoate,metrizoate, ioxaglate), barium sulfate, thorium dioxide, gold, goldnanoparticle, gold nanoparticle aggregate, fluorophore, two-photonfluorophore, or a hapten. In embodiments, the chain terminator ortransfer agent includes an ethyl vinyl ether. In embodiments, the chainterminator or transfer agent includes an alkene containing substitutedor unsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl or substituted orunsubstituted heteroaryl. In embodiments, the chain terminator ortransfer agent includes an alkene bonded to a substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl or substituted orunsubstituted heteroaryl. In embodiments, the chain terminator ortransfer agent includes an alkene containing compound (e.g., alsoincluding a function group, substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, substituted or unsubstituted heteroaryl, ordetectable moiety). In embodiments, the chain terminator or transferagent includes a moiety selected from:

In embodiments the chain terminator or transfer agent is a solidsupport. In embodiments, the chain terminator or transfer agent is ananoparticle. In embodiments, the chain terminator or transfer agent isa substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl orsubstituted or unsubstituted heteroaryl. In embodiments, the chainterminator or transfer agent is a functional moiety. In embodiments, thechain terminator or transfer agent is a detectable moiety. Inembodiments, the chain terminator or transfer agent includes a ³²P,fluorescent dye, electron-dense reagent, enzyme (e.g., as commonly usedin an ELISA), biotin, digoxigenin, paramagnetic molecule, paramagneticnanoparticle, ultrasmall superparamagnetic iron oxide (“USPIO”)nanoparticles, USPIO nanoparticle aggregate, superparamagnetic ironoxide (“SPIO”) nanoparticle, SPIO nanoparticle aggregate,monochrystalline SPIO, monochrystalline SPIO aggregate, monochrystallineiron oxide nanoparticle, monochrystalline iron oxide, other nanoparticlecontrast agent, liposome or other delivery vehicle containing Gadoliniumchelate (“Gd-chelate”) molecule, Gadolinium, radioisotope, radionuclide(e.g. carbon-11, nitrogen-13, oxygen-15, fluorine-18, rubidium-82),fluorodeoxyglucose (e.g. fluorine-18 labeled), any gamma ray emittingradionuclids, positron-emitting radionuclide, radiolabeled glucose,radiolabeled water, radiolabeled ammonia, biocolloids, microbubble (e.g.including microbubble shell including albumin, galactose, lipid, and/orpolymers; microbubble gas core including air, heavy gas(es),perfluorcarbon, nitrogen, octafluoropropane, perflexane lipidmicrosphere, perflutren, etc.), iodinated contrast agent (e.g. iohexol,iodixanol, ioversol, iopamidol, ioxilan, iopromide, diatrizoate,metrizoate, ioxaglate), barium sulfate, thorium dioxide, gold, goldnanoparticle, gold nanoparticle aggregate, fluorophore, two-photonfluorophore, or a hapten. In embodiments, the chain terminator ortransfer agent is an ethyl vinyl ether. In embodiments, the chainterminator or transfer agent is an alkene containing substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl or substituted orunsubstituted heteroaryl. In embodiments, the chain terminator ortransfer agent is an alkene bonded to a substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl or substituted or unsubstitutedheteroaryl. In embodiments, the chain terminator or transfer agent is analkene containing compound (e.g., also including a function group,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl,substituted or unsubstituted heteroaryl, or detectable moiety).

In embodiments, the method includes radical polymerization, controlledradical polymerization, reversible addition-fragmentation chain transfer(RAFT) polymerization, atom transfer radical polymerization (ATRP),ring-opening metathesis polymerization (ROMP), anionic and cationicpolymerizations, free radical living polymerization, acyclic dienemetathesis polymerization, radiation-induced polymerization,ring-opening olefin metathesis polymerization, polycondensationreactions, or iniferter-induced polymerization. In embodiments, themethod employs radical polymerization, controlled radicalpolymerization, reversible addition-fragmentation chain transfer (RAFT)polymerization, atom transfer radical polymerization (ATRP),ring-opening metathesis polymerization (ROMP), anionic and cationicpolymerizations, free radical living polymerization, acyclic dienemetathesis polymerization, radiation-induced polymerization,ring-opening olefin metathesis polymerization, polycondensationreactions, or iniferter-induced polymerization.

In embodiments, the method includes ring-opening metathesispolymerization. In embodiments, the method employs ring-openingmetathesis polymerization.

In an aspect is provided a method of making an amphiphilic block graftcopolymer, the method including: (i) reacting a plurality ofoligonucleotide monomers with a polymerization catalyst thereby forminga hydrophilic block portion, wherein each of the plurality ofoligonucleotide monomers includes a polymerizable monomer covalentlybound to an oligonucleotide; (ii) reacting the hydrophilic block portionwith a plurality of hydrophobic monomers and the polymerization catalystthereby forming the amphiphilic block graft copolymer, wherein each ofthe plurality of hydrophobic monomers includes the polymerizable monomercovalently bound to a hydrophobic moiety.

In embodiments, the amphiphilic block graft copolymer, oligonucleotidemonomers, polymerization catalyst, polymerizable monomer,oligonucleotide, of hydrophobic monomers, and hydrophobic moiety, are asdescribed herein, including in aspects, embodiments, examples, figures,tables, schemes, and claims.

In an aspect is provided a method of making an amphiphilic block graftcopolymer, the method including: (i) reacting a plurality of hydrophobicmonomers with a polymerization catalyst thereby forming a hydrophobicblock portion, wherein each of the plurality of hydrophobic monomersincludes a polymerizable monomer covalently bound to a hydrophobicmoiety; (ii) reacting the hydrophobic block portion with a plurality ofoligonucleotide monomers and the polymerization catalyst thereby formingthe amphiphilic block graft copolymer, wherein each of the plurality ofoligonucleotide monomers includes the polymerizable monomer covalentlybound to an oligonucleotide.

In embodiments, the amphiphilic block graft copolymer, oligonucleotidemonomers, polymerization catalyst, polymerizable monomer,oligonucleotide, of hydrophobic monomers, and hydrophobic moiety, are asdescribed herein, including in aspects, embodiments, examples, figures,tables, schemes, and claims.

In embodiments, the method includes radical polymerization, controlledradical polymerization, reversible addition-fragmentation chain transfer(RAFT) polymerization, atom transfer radical polymerization (ATRP),ring-opening metathesis polymerization (ROMP), anionic and cationicpolymerizations, free radical living polymerization, acyclic dienemetathesis polymerization, radiation-induced polymerization,ring-opening olefin metathesis polymerization, polycondensationreactions, or iniferter-induced polymerization. In embodiments, themethod employs radical polymerization, controlled radicalpolymerization, reversible addition-fragmentation chain transfer (RAFT)polymerization, atom transfer radical polymerization (ATRP),ring-opening metathesis polymerization (ROMP), anionic and cationicpolymerizations, free radical living polymerization, acyclic dienemetathesis polymerization, radiation-induced polymerization,ring-opening olefin metathesis polymerization, polycondensationreactions, or iniferter-induced polymerization.

In embodiments, the method includes ring-opening metathesispolymerization. In embodiments, the method employs ring-openingmetathesis polymerization.

In embodiments, the method is a method described herein, including in anaspect, embodiment, example, figure, table, scheme, and claim.

IV. Methods of Using Polymers

In an aspect is provided a method of internalize nucleic acids (e.g.,nucleic acids included in the graft polymer) into a cell includingcontacting the cell with a graft polymer. In embodiments, the graftpolymer is described herein, including in an aspect, embodiment,example, figure, table, scheme, or claim. In embodiments, the nucleicacid is an oligonucleotide as described herein.

In an aspect is provided a method of regulating an mRNA level in a cellincluding contacting the cell with a graft polymer (e.g., wherein thegraft polymer includes nucleic acids capable of regulating an mRNAlevel). In embodiments, the graft polymer is described herein, includingin an aspect, embodiment, example, figure, table, scheme, or claim. Inembodiments, regulating is reducing. In embodiments, the method includescontacting the cell with the graft polymer. In embodiments, the nucleicacid included in the graft polymer is complementary to a sequence of themRNA. In embodiments, the nucleic acid included in the graft polymerhybridizes to the mRNA. In embodiments, the nucleic acid included in thegraft polymer selectively hybridizes to the mRNA. In embodiments, themethod includes gene interference (e.g., by the nucleic acid). Inembodiments, the nucleic acid is an oligonucleotide as described herein.

In an aspect is provided a method of detecting a first nucleic acid in acell including contacting the cell with a graft polymer (e.g., whereinthe graft polymer includes a second nucleic acid capable of hybridizingto the first nucleic acid (e.g., selectively hybridizing)). Inembodiments, the graft polymer is described herein, including in anaspect, embodiment, example, figure, table, scheme, or claim. Inembodiments, the second nucleic acid included in the graft polymer iscomplementary to a sequence of the first nucleic acid (e.g., DNA orRNA). In embodiments, the second nucleic acid is an oligonucleotide asdescribed herein. In embodiments, the graft polymer includes adetectable moiety.

In an aspect is provided a method of detecting a DNA sequence in a cellincluding contacting the cell with a graft polymer (e.g., wherein thegraft polymer includes a nucleic acid capable of hybridizing to the DNAsequence (e.g., selectively hybridizing)). In embodiments, the graftpolymer is described herein, including in an aspect, embodiment,example, figure, table, scheme, or claim. In embodiments, the nucleicacid included in the graft polymer is complementary to a portion of theDNA sequence. In embodiments, the nucleic acid is an oligonucleotide asdescribed herein. In embodiments, the graft polymer includes adetectable moiety.

In an aspect is provided a method of detecting a RNA sequence in a cellincluding contacting the cell with a graft polymer (e.g., wherein thegraft polymer includes a nucleic acid capable of hybridizing to the RNAsequence (e.g., selectively hybridizing)). In embodiments, the graftpolymer is described herein, including in an aspect, embodiment,example, figure, table, scheme, or claim. In embodiments, the nucleicacid included in the graft polymer is complementary to a portion of theRNA sequence. In embodiments, the nucleic acid is an oligonucleotide asdescribed herein. In embodiments, the graft polymer includes adetectable moiety.

In an aspect is provided a method of purifying a first nucleic acidincluding contacting the first nucleic acid with a graft polymer (e.g.,wherein the graft polymer includes a second nucleic acid capable ofhybridizing to the first nucleic acid (e.g., selectively hybridizing)).In embodiments, the graft polymer is described herein, including in anaspect, embodiment, example, figure, table, scheme, or claim. Inembodiments, the second nucleic acid included in the graft polymer iscomplementary to a sequence of the first nucleic acid (e.g., DNA orRNA). In embodiments, the second nucleic acid is an oligonucleotide asdescribed herein. In embodiments, the graft polymer includes adetectable moiety.

In an aspect is provided a method of administering a nucleic acid to acell including contacting the cell with a graft polymer, wherein thegraft polymer includes the nucleic acid. In embodiments, the graftpolymer is described herein, including in an aspect, embodiment,example, figure, table, scheme, or claim. In embodiments, the nucleicacid is an oligonucleotide as described herein.

In an aspect is provided a method of administering a first nucleic acidto a cell including contacting the cell with a graft polymer (e.g.,wherein the graft polymer includes a second nucleic acid hybridized tothe first nucleic acid (e.g., selectively hybridizing)). In embodiments,the graft polymer is described herein, including in an aspect,embodiment, example, figure, table, scheme, or claim. In embodiments,the second nucleic acid included in the graft polymer is complementaryto a sequence of the first nucleic acid (e.g., DNA or RNA). Inembodiments, the second nucleic acid is an oligonucleotide as describedherein.

In embodiments, the graft polymer is a graft polymer described herein(including in an aspect, embodiment, example, figure, table, claim, orscheme). In embodiments, the graft polymer is a brush polymer. Inembodiments, the graft polymer is a block graft copolymer. Inembodiments, the graft polymer is a block graft copolymer describedherein, including in an aspect, embodiment, example, figure, table,scheme, or claim.

V. Additional Embodiments

1. A graft polymer comprising a linear backbone covalently bound to aplurality of oligonucleotide branches, wherein said graft polymer isassembled by graft-through polymerization of a plurality ofoligonucleotide monomers comprising a polymerizable monomer covalentlybound to an oligonucleotide, said oligonucleotide thereby forming eachof said plurality of oligonucleotide branches.2. The graft polymer of embodiment 1, wherein said oligonucleotidecomprises at least 3 nucleobases and at least 2 different nucleobases.3. The graft polymer of embodiment 1, wherein said oligonucleotidecomprises at least 5 nucleobases and at least 3 different nucleobases.4. The graft polymer of embodiment 1, wherein said oligonucleotidecomprises at least 10 nucleobases and at least 4 different nucleobases.5. The graft polymer of embodiment 1 comprising at least 3oligonucleotide branches.6. The graft polymer of embodiment 1 comprising at least 5oligonucleotide branches.7. The graft polymer of embodiment 1 comprising at least 10oligonucleotide branches.8. The graft polymer of embodiment 1 having the formula:R¹-[M(O)]_(n)—R² wherein, n is an integer from 2 to 1000; M is thepolymerized product of the polymerizable monomer; O is theoligonucleotide; and R1 and R2 are terminal polymer moieties.9. The graft polymer of embodiment 1, wherein the graft-throughpolymerization employs ring-opening metathesis polymerization (ROMP).10. The graft polymer of embodiment 1, wherein the linear backbone is apolynorbornyl chain.11. A method of making a graft polymer, the method comprising:(i) reacting a plurality of oligonucleotide monomers with apolymerization catalyst or initiator, wherein each of said plurality ofoligonucleotide monomers comprises a polymerizable monomer covalentlybound to an oligonucleotide; and(ii) terminating said reacting with a chain terminator or transferagent.12. The method of embodiment 11, wherein the graft polymer comprising alinear backbone covalently bound to a plurality of oligonucleotidebranches.13. The method of embodiment 11, wherein the oligonucleotide comprisesat least 3 nucleobases and at least 2 different nucleobases.14. The method of embodiment 11, wherein the oligonucleotide comprisesat least 5 nucleobases and at least 3 different nucleobases.15. The method of embodiment 11, wherein the oligonucleotide comprisesat least 10 nucleobases and at least 4 different nucleobases.16. The method of embodiment 11, wherein the polymer comprises at least3 oligonucleotide branches.17. The method of embodiment 11, wherein the polymer comprises at least5 oligonucleotide branches.18. The method of embodiment 11, wherein the polymer comprises at least10 oligonucleotide branches.19. The method of embodiment 11, wherein the polymer comprises a linearbackbone comprising a polynorbornyl chain.20. The method of embodiment 11, wherein the polymer has the formula:R¹-[M(O)]_(n)—R² wherein, n is an integer from 2 to 1000; M is thepolymerized product of the polymerizable monomer; O is theoligonucleotide; and R¹ and R² are terminal polymer moieties.21. The method of one of embodiments 11 to 20, comprising radicalpolymerization, controlled radical polymerization, reversibleaddition-fragmentation chain transfer (RAFT) polymerization, atomtransfer radical polymerization (ATRP), ring-opening metathesispolymerization (ROMP), anionic and cationic polymerizations, freeradical living polymerization, acyclic diene metathesis polymerization,radiation-induced polymerization, ring-opening olefin metathesispolymerization, polycondensation reactions, or iniferter-inducedpolymerization.22. The method of one of embodiments 11 to 20, comprising ring-openingmetathesis polymerization.23. An amphiphilic block graft copolymer comprising a linear backbonecovalently bound to a plurality of oligonucleotide branches and aplurality of hydrophobic side chains, wherein: said plurality ofoligonucleotide branches form a hydrophilic block portion of saidamphiphilic graft copolymer and said hydrophobic side chains form ahydrophobic block portion of said amphiphilic graft copolymer; saidgraft copolymer is assembled by graft-through polymerization of aplurality of oligonucleotide monomers and a plurality of hydrophobicmonomers, wherein each of said plurality of oligonucleotide monomerscomprises a polymerizable monomer covalently bound to anoligonucleotide, said oligonucleotide thereby forming each of saidplurality of oligonucleotide branches; and each of said plurality ofhydrophobic monomers comprises said polymerizable monomer covalentlybound to a hydrophobic moiety, said hydrophobic moiety thereby formingeach of said plurality of hydrophobic side chains.24. The amphiphilic block graft copolymer of embodiment 23, wherein saidoligonucleotide comprises at least 3 nucleobases and at least 2different nucleobases.25. The amphiphilic block graft copolymer of embodiment 23, wherein saidoligonucleotide comprises at least 5 nucleobases and at least 3different nucleobases.26. The amphiphilic block graft copolymer of embodiment 23, wherein saidoligonucleotide comprises at least 10 nucleobases and at least 4different nucleobases.27. The amphiphilic block graft copolymer of embodiment 23 comprising atleast 3 oligonucleotide branches.28. The amphiphilic block graft copolymer of embodiment 23 comprising atleast 5 oligonucleotide branches.29. The amphiphilic block graft copolymer of embodiment 23 comprising atleast 10 oligonucleotide branches.30. The amphiphilic block graft copolymer of one of embodiments 23 to29, wherein said hydrophobic moiety is a substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, or substituted or unsubstitutedheteroaryl.31. The amphiphilic block graft copolymer of one of embodiments 23 to29, wherein said hydrophobic moiety is an unsubstituted benzyl.32. The amphiphilic block graft copolymer of embodiment 23 having theformula: R¹-[M(O)]_(n)-[M(P)]_(m)—R² or R¹-[M(P)]_(m)-[M(O)]_(n)—R²wherein, n is an integer from 2 to 1000; m is an integer from 2 to 1000;M is the polymerized product of the polymerizable monomer; O is theoligonucleotide; P is the hydrophobic moiety; and R¹ and R² are terminalpolymer moieties.33. A micelle comprising the amphiphilic block graft copolymer of one ofembodiments 23 to 32.34. The micelle of embodiment 33, having a diameter of between about 1and about 1000 nm.35. The micelle of embodiment 33, having a diameter of between about 5and about 100 nm.36. The micelle of embodiment 33, having a diameter of between about 10and about 50 nm.37. A nanoparticle comprising the amphiphilic block graft copolymer ofone of embodiments 23 to 32.38. The nanoparticle of embodiment 37, wherein said nanoparticle is aspherical nanoparticle.39. The nanoparticle of one of embodiments 37 to 38, having a diameterof between about 1 and about 1000 nm.40. The nanoparticle of one of embodiments 37 to 38, having a diameterof between about 5 and about 100 nm.41. The nanoparticle of one of embodiments 37 to 38, having a diameterof between about 10 and about 50 nm.42. A method of making an amphiphilic block graft copolymer, the methodcomprising:(i) reacting a plurality of oligonucleotide monomers with apolymerization catalyst thereby forming a hydrophilic block portion,wherein each of said plurality of oligonucleotide monomers comprises apolymerizable monomer covalently bound to an oligonucleotide;(ii) reacting said hydrophilic block portion with a plurality ofhydrophobic monomers and said polymerization catalyst thereby formingsaid amphiphilic block graft copolymer, wherein each of said pluralityof hydrophobic monomers comprises said polymerizable monomer covalentlybound to a hydrophobic moiety.43. The method of embodiment 42, wherein the oligonucleotide comprisesat least 3 nucleobases and at least 2 different nucleobases.44. The method of embodiment 42, wherein the oligonucleotide comprisesat least 5 nucleobases and at least 3 different nucleobases.45. The method of embodiment 42, wherein the oligonucleotide comprisesat least 10 nucleobases and at least 4 different nucleobases.46. The method of embodiment 42, wherein the amphiphilic block graftcopolymer comprises at least 3 oligonucleotide branches.47. The method of embodiment 42, wherein the amphiphilic block graftcopolymer comprises at least 5 oligonucleotide branches.48. The method of embodiment 42, wherein the amphiphilic block graftcopolymer comprises at least 10 oligonucleotide branches.49. The method of embodiment 42, wherein the amphiphilic block graftcopolymer comprises a linear backbone comprising a polynorbornyl chain.50. The method of embodiment 42, wherein the amphiphilic block graftcopolymer of embodiment 23 has the formula: R¹-[M(O)]_(n)-[M(P)]_(m)—R²wherein, n is an integer from 2 to 1000; m is an integer from 2 to 1000;M is the polymerized product of the polymerizable monomer; O is theoligonucleotide; P is the hydrophobic moiety; and R¹ and R² are terminalpolymer moieties.51. The method of one of embodiments 42 to 50, comprising atom transferradical polymerization (ATRP), ring-opening metathesis polymerization(ROMP), anionic and cationic polymerizations, free radical livingpolymerization, radiation-induced polymerization, ring-opening olefinmetathesis polymerization, polycondensation reactions, oriniferter-induced polymerization.52. The method of one of embodiments 42 to 50, comprising ring-openingmetathesis polymerization.53. A method of making an amphiphilic block graft copolymer, the methodcomprising:(i) reacting a plurality of hydrophobic monomers with a polymerizationcatalyst thereby forming a hydrophobic block portion, wherein each ofsaid plurality of hydrophobic monomers comprises a polymerizable monomercovalently bound to a hydrophobic moiety;(ii) reacting said hydrophobic block portion with a plurality ofoligonucleotide monomers and said polymerization catalyst therebyforming said amphiphilic block graft copolymer, wherein each of saidplurality of oligonucleotide monomers comprises said polymerizablemonomer covalently bound to an oligonucleotide.54. The method of embodiment 53, wherein the oligonucleotide comprisesat least 3 nucleobases and at least 2 different nucleobases.55. The method of embodiment 53, wherein the oligonucleotide comprisesat least 5 nucleobases and at least 3 different nucleobases.56. The method of embodiment 53, wherein the oligonucleotide comprisesat least 10 nucleobases and at least 4 different nucleobases.57. The method of embodiment 53, wherein the amphiphilic block graftcopolymer comprises at least 3 oligonucleotide branches.58. The method of embodiment 53, wherein the amphiphilic block graftcopolymer comprises at least 5 oligonucleotide branches.59. The method of embodiment 53, wherein the amphiphilic block graftcopolymer comprises at least 10 oligonucleotide branches.60. The method of embodiment 53, wherein the amphiphilic block graftcopolymer comprises a linear backbone comprising a polynorbornyl chain.61. The method of embodiment 53, wherein the amphiphilic block graftcopolymer of embodiment 23 has the formula: R¹-[M(P)]_(m)-[M(O)]_(n)—R²wherein, n is an integer from 2 to 1000; m is an integer from 2 to 1000;M is the polymerized product of the polymerizable monomer; O is theoligonucleotide; P is the hydrophobic moiety; and R¹ and R² are terminalpolymer moieties.62. The method of one of embodiments 53 to 61, comprising atom transferradical polymerization (ATRP), ring-opening metathesis polymerization(ROMP), anionic and cationic polymerizations, free radical livingpolymerization, radiation-induced polymerization, ring-opening olefinmetathesis polymerization, polycondensation reactions, oriniferter-induced polymerization.63. The method of one of embodiments 53 to 61, comprising ring-openingmetathesis polymerization.

VI. Examples

The following examples are offered to illustrate, but not to limit, theclaimed invention.

Example 1: Methods of Making Poly(Oligonucleotides)

This example describes an efficient synthetic strategy for theincorporation of nucleic acids into particle and polymer-basedmaterials. This work is published in James et al.,“Poly(oligonucleotide),” J Am Chem Soc, in press, and Thompson et al.,“Labelling Polymers and Micellar Nanoparticles via Initiation,Propagation and Termination with ROMP,” Polym Chem, 2014 Mar. 21;5(6):1954-1964, the contents of which are hereby incorporated in theirentirety for all purposes.

Here we report the preparation of poly(oligonucleotide) brush polymersand amphiphilic brush copolymers from nucleic acid monomers viagraft-through polymerization. We describe the polymerization ofPNA-norbornyl monomers to yield poly-PNA (poly(peptide nucleic acid))via ing-opening metathesis polymerization (ROMP) with the initiator,(IMesH₂)—(C₅H₅N)₂(Cl)₂RuCHPh.¹ In addition, we present the preparationof poly-PNA nanoparticles from amphiphilic block copolymers and describetheir hybridization to a complementary single-stranded DNA (ssDNA)oligonucleotide.

The display of chemical functionality in a multivalent fashion onsurfaces and particles and as brushes on polymer backbones is a commontheme in nature as well as for synthetic systems.²⁻⁴ Such systems takeadvantage of the unique properties that arise when monomeric species areincorporated into a densely packed three-dimensional (3D) architecture.Here we describe the preparation of polymeric nucleic acids whereinsingle-stranded sequences of peptide nucleic acids (PNAs) areincorporated as polymer brushes via graft-through polymerization usingthe ROMP initiator (IMesH₂)—(C₅H₅N)₂(Cl)₂RuCHPh (FIGS. 1A-1C). Nucleicacids, both natural and synthetic, standout as the quintessentialcarriers of chemical information stored as specific sequences of basespositioned along a backbone.⁴⁻⁷ As such, synthetic oligonucleotides andnucleic acid bioconjugates are powerful tools in a range of fieldsincluding in biotechnology (e.g., PCR)^(8,9) and in materials science asprogrammable structural synthons¹⁰⁻¹⁶ and as aptamers selected by invitro evolution.¹⁷⁻²¹ In each application the nucleic acid functions toenrich a chemical system with information, facilitating predictableinteractions with complementary sequences,²² or with other moleculesincluding enzymes, proteins, and small molecules.^(18, 23-25) Wereasoned that an approach allowing the graft-through polymerization ofan oligonucleotide sequence would provide a powerful new tool for themultivalent display of chemical information on a synthetic template.

There have been extensive efforts to prepare nucleic acid inspiredsynthetic polymers involving the direct polymerization of appropriatelymodified monomers, generating synthetic polymers with single nucleobasesas side-chains.²⁶⁻³¹ Although this approach allows the integration ofpurine and pyrimidine bases onto a synthetic backbone, it does not allowthe incorporation of sequences containing multiple bases and thus doesnot result in informational polymeric systems. In addition there are anincreasing number of examples of oligonucleotide-polymer bioconjugatesin the literature, each reliant upon post-polymerization conjugationreactions.³²⁻³⁴ These approaches, shown in Scheme 1, seek to fixrecognition elements natural to DNA and RNA along a synthetic polymer orpolymeric nanoparticle template and have found use in an array of arenasincluding the programmed assembly of nanoparticles,^(2,35-37) indelivery vehicles,^(5,38-40) and as effective DNA-probes.^(34,41,42)Furthermore, the function of these materials is intrinsically governedby the information within the nucleic acid sequence itself as well asthe dense and multivalent 3D array induced by the polymer scaffold.Indeed, function dictated by 3D biomolecular display is not unique tonucleic acids, rather this concept extends to all classes ofbiomolecules, most effectively demonstrated in the past with peptidesand proteins.^(3,43-47) Strategies for the polymerization of(graft-through) and polymerization from (graft-from) proteins andpeptides have been used to build macromolecules through sequentialaddition of monomers to a growing chain, taking advantage ofpolymerization catalyst proficiency and avoiding kinetically unfavorableconjugations (graft-to) between multiple large macromolecules.⁴⁸⁻⁵⁴However, unlike for other bio-molecules (saccharides,^(55,57)peptides,⁵⁸ and proteins^(59,60)), there are no examples ofgraft-through polymerization and few examples of graft-frompolymerization of nucleic acids.^(61,62) Therefore, despite theirpromise, polymer bioconjugates of true nucleic acid sequences have beenmostly limited to those prepared via post-polymerization modificationand hence are difficult to reproduce and suffer from incompleteincorporation of the nucleic acid at each position of the polymer (FIGS.4A-4B).

In order to avoid shortcomings associated with post-polymerizationmodification reactions, a nucleic acid monomer capable of undergoingdirect graft-through polymerization was synthesized. Initial studiesattempting direct-polymerization of DNA-based monomers via ROMP were metwith limited success. Therefore, a PNA-based monomer was chosen as anideal target for this study for three key reasons: (1) PNA can beprepared in milligram to gram quantities via standard peptidebond-forming reactions on solid support; (2) PNA is soluble in DMFmaking it readily compatible with (IMesH₂)—(C₅H₅N)₂(Cl)₂RuCHPh (FIGS.1A-1C for structure); and (3) we hypothesized that the neutralN-(2-amino-ethyl)-glycine back-bone would be more compatible with theruthenium-based catalyst than the polyanionic phosphate backbone of DNA.The 10-base PNA sequence (FIGS. 1A-1C) was designed to discourageself-hybridization, while providing a sufficient number of bases forefficient hybridization with a comple-mentary sequence of DNA at roomtemperature. Moreover, this 10-base sequence represents a sequenceencoded with each of the four letters of the genetic alphabet and enoughinformation to communicate specifically with other nucleic acids andproteins, a function not possible via the polymer Polymerization of asingle purine of pyrimidine monomer (as in FIG. 4B). After the completePNA sequence was prepared,N-(glycine)-cis-5-norbornene-exo-dicarboximide was coupled to theN-terminus while on solid support. The PNA norbornene monomer (PNA-Nb)was then cleaved and deprotected from the solid support using TFA:cresol(80:20), purified by HPLC, and the mass confirmed by ESI-MS. FollowingHPLC purification, PNA-Nb was lyophilized to afford a white powder.

For polymerization studies, PNA-Nb was resuspended in dry, degassedN,N-dimethylformamide-d₇ in a J. Young NMR tube in a glovebox. Anappropriate amount of ruthenium initiator was added to the solution, andthe disappearance of the norbornene olefin resonance was then monitoredby ¹H NMR. Complete disappearance of the monomeric olefin peak indicatedcomplete polymerization of PNA-Nb into poly-PNA. A series of experimentswere carried out to determine reproducibility of polymerizationreactions with respect to both the preparation of homopolymers as wellas block copolymers (Table 1).

TABLE 1 Polymers and Copolymers of PNA with Monomers Shown in FIG. 2.polymer mon₁ ^(a) mon₂ ^(c) m^(d) n^(d) % con.^(e) I PNA-Nb (10:1)^(b) —10 — 99 II 1 (35:1) PNA-Nb (5:1) 35 5 97 III PNA-Nb (5:1) — 5 — 97 IV 1(30:1) PNA-Nb (7.5:1) 30 5 65 V 1 (30:1) PNA-Nb (7.5:1) 30 5 65 VI 1(30:1) PNA-Nb (7.5:1) 30 6 79 VII 1 (36:1) PNA-Nb (9:1) 35 8 88 VIII 1(36:1) PNA-Nb (18:1) 35 16  87 IX 2 (36:1) PNA-Nb (9:1) 33 7 74 X 3(36:1) PNA-Nb (9:1) 41 5 56 ^(a)Indicates indentity of monomerpolymerized first (degree of polymerization, DP = m). ^(b)Ratios shownindicate monomer to initiator ratio or intended DP. ^(c)Indicatesidentity of monomer polymerized second (DP = n). ^(d)Observed degree ofpolymerization of mon₁ (m) or mon₂ (n). ^(e)Percent conversion of PNA-Nbdetermined by ¹H NMR.

For PNA homopolymers, degrees of polymerization of 5 (polymer I) and 10(polymer III) were targeted with complete consumption of PNA-Nbconfirmed by ¹H NMR for both reactions. In preparation of aPNA-containing block copolymer, a phenyl-functionalized norbornene (1)was polymerized as the first block followed by PNA-Nb incorporated asthe second block (as shown in polymer II (FIGS. 1A-1C, Table 1). Toachieve this, PNA-Nb was added to the living phenyl polymer chain, andthe disappearance of the norbornene olefin of PNA-Nb was monitored by ¹HNMR. SEC-MALS confirmed molecular weights for the resulting species.Having established that PNA-Nb could be successfully polymerized intoblock copolymers, we sought to assess the reproducibility of thesereactions by attempting to synthesize block copolymers of identicalcomposition. Using a live ruthenium catalyst on a phenyl homopolymerwith a degree of polymerization of 30, three separate but identicalreactions were set up in which the attempted degree of polymerization ofthe PNA monomer was 7.5 (Table 1, polymers IV-VI). The degree ofpolymerization of the PNA block ranged from 5 to 6 (60-80% completion),indicating a good degree of reproducibility and predictability for thesereactions. In addition, higher degrees of polymerization could beachieved for this type of block copolymer as illustrated by thepreparation of polymers VII and VIII (Table 1). To examine thecompatibility of PNA-Nb polymerization with other block copolymersystems, an oligoethylene glycol functionalized norbornene (2) and aquaternary amine-functionalized norbornene (3) were synthe-sized asmonomers for incorporation into block copolymer scaffolds as the initialblock. The resulting block polymers (IX and X in Table 1) showed percentconversions of PNA-Nb comparable to the phenyl-based block copolymers,with the amine-functionalized system demonstrating the lowest percentconversion. Given the slight variation in PNA-Nb percent conversionbetween these three different block copolymer systems (VII, IX and X inTable 1), the identity of the non-PNA block may dictate PNA conversionefficiency and should be taken into consideration for future studies. Toassess the DNA-binding capability of these systems, block copolymer IIwas chosen. The assembly of II to generate spherical nanoparticles wasachieved by dissolving in DMSO and then dialyzing into aqueoussolution.⁶³ The resulting nanoparticle (PNA-NP) was characterized by DLSand TEM (FIGS. 3A-3E). DLS data support the formation of an aggregatedspecies in solution. TEM reveals nanoparticles on the order of 20 nm indiameter. The melting temperature (T_(m)) of PNA-NP hybridized with itscomplementary DNA sequence was determined to be 58.1° C., an ˜8° C.increase over an identical, nonparticulate, PNA sequence. These meltingdata suggest cooperative binding and accessible PNA forming the shell ofthe nanoparticles. In support of this model, we conducted a moleculardynamics simulation of PNA-NP⁶⁴⁻⁶⁶ assembled from 60 amphiphiles givinga structure that equilibrated into a spherical particle ˜21 nm indiameter. Polynorbornyl chains packed well to form a compact hydrophobiccore largely protected from contact with water. The simulations show thehydrophilic PNA chains solvated in water forming the shell of themicelles.

In summary, we have shown that one can prepare nucleic acid brushpolymers and amphiphilic brush copolymers by direct polymerization viagraft-through polymerization of a nucleic acid. To our knowledge, thisis the first example of a polymer-nucleic acid bioconjugate generatedvia direct polymerization of an oligonucleotide monomer. In addition,these materials show cooperative hybridization to complementary DNAoligonucleotides. We believe this type of approach provides an efficientsynthetic strategy for the incorporation of nucleic acids into particleand polymer-based materials. The interest in doing so is driven bypotential applications including the facile preparation of materials foraffinity purification of DNA,^(67,68) gene and nucleic acid delivery tocells,^(5,38-40,69-72) and in the development of materials capable ofprogrammed self-assembly.^(12-16,73-77)

1. General Methods

All reagents were purchased from commercial sources and used withoutfurther purification unless otherwise indicated.N-phenyl-cis-5-norbornene-exo-dicarboximide [Ku, T. H.; Chien, M. P.;Thompson, M. P.; Sinkovits, R. S.; Olson, N. H.; Baker, T. S.;Gianneschi, N. C. J. Am. Chem. Soc., 2011, 133, 8392],2-(2,5,8,11-tetraoxatridecan-13-yl)-3a,4,7,7a-tetrahydro-1H-4,7-methanoisoindole-1,3(2H)-dione[Thompson, M. P.; Randolph, L. M.; James, C. R.; Davalos, A. N.; Hahn,M. E.; Gianneschi, N. C. Polym. Chem., 2014, 5, 1954],N-benzyl-2-(1,3-dioxo-1,3,3a,4,7,7a-hexahydro-2H-4,7-epoxyisoindol-2-yl)-N,N-dimethylethan-1-aminium[Rankin, D. A.; P'Pool, S. J.; Schanz, H.-J.; Lowe, A. B. J. Polym. Sci.A Polym. Chem., 2007, 45, 2113],(peg)N-(glycine)-cis-5-norbornene-exo-dicarboximide [Conrad, R. M.;Grubbs, R. H. Angew. Chem. Int. Edit., 2009, 48, 8328] were prepared asdescribed. All PNA sequences were made using NovaPEG Rink Amide resin,purchased from EMD Millipore, with a loading of 0.49 mmol/g and aswelling volume of 8.8 mL/g in CH²Cl². The synthesis of all PNAoligomers used Fmoc/Bhoc protected monomers (Fmoc-A(Bhoc)-aeg-OH,Fmoc-C(Bhoc)-aeg-OH, Fmoc-G(Bhoc)-aeg-OH, and Fmoc-T-aeg-OH) purchasedfrom Panagene. PNA sequences were manually synthesized in house usingstandard solid-phase peptide synthesis conditions. RP-HPLC analysis ofPNAs was performed on a Hitachi-Elite LaChrom L-2130 pump with astep-wise gradient. Detection was at 260 nm using an in-line UV-Visdetector (Hitachi-Elite LaChrom L-2420). For analysis, an analyticalscale Phenomenex Jupiter 4 μm Proteo 90 Å column (150×4.60 mm) wasutilized. For purification, a semi-preparative Phenomenex Jupiter 4 μmProteo 90 Å column (250×10.0 mm) was utilized. Mass spectra wereobtained at the UCSD Chemistry and Biochemistry Molecular MassSpectrometry Facility. Low-resolution mass spectra were obtained using aThermo LCQdeca mass spectrometer. Concentrations of oligonucleotides andpeptide nucleic acids were determined using a Thermo Scientific NanoDrop2000c spectrophotometer. Modified 2^(nd) Generation Grubbs' Rutheniuminitiator (IMesH₂)(C₅H₅N)₂(Cl)₂Ru═CHPh was prepared as described bySanford et. al. [Sanford, M. S.; Love, J. A.; Grubbs, R. H.Organometallics, 2001, 20, 5314] Sealed ampules of (CD₃)₂NCOD (DMF-d₇)used in polymerization reactions was purchased from Cambridge IsotopeLaboratories Inc. and was distilled and degassed with 3 freeze-pump-thawcycles prior to use. ¹H (400 MHz) NMR spectra were recorded on a VarianMercury Plus spectrometer. Chemical shifts (¹H) are reported in δ (ppm)relative to the DMF-d₇ residual proton peaks (8.03, 2.92, and 2.75 ppm).Polymer molecular weight and dispersity were determined viasize-exclusion chromatography (Phenomenex Phenogel 5 μm 10, 1K-75K,300×7.80 mm in series with a Phenomex Phenogel 5 μm 10, 10K-1000K,300×7.80 mm (mobile phase: 0.05 M LiBr in DMF)) using a Hitachi-EliteLaChrom L-2130 pump equipped with a DAWN HELEOS multi-angle lightscattering (MALS) detector (Wyatt Technology) and a refractive indexdetector (Hitachi L-2490) normalized to a 30,000 g/mol polystyrenestandard. The dn/dc values used were 0.179. Hydrodynamic diameter(D_(h)) of nanoparticles was measured via DLS using a DynaPro NanoStar(Wyatt Technology). TEM samples were deposited on carbon/formvar-coatedcopper grids (Ted Pella Inc.), stained with 1% w/w uranyl acetate, andimaged using a Technai G2 Sphera operating at an accelerating voltage of200 kV. Complementary and non-complementary DNA sequences were purchasedfrom Integrated DNA Technologies (purified by HPLC, confirmed byESI-MS). DNA melting temperature analysis was conducted using a CarySeries 100 UV-Vis spectrophotometer equipped with a Cary temperaturecontroller.

2. PNA Monomer Synthesis

All PNA sequences were manually synthesized. All reactions and washes ofthe resin were performed in a fritted glass peptide synthesis vessel,with the exception of the cleavage and deprotection of the oligomer fromthe resin, which was done in a polypropylene Poly-Prep ChromatographyColumn, purchased from Bio-Rad Laboratories. Unless otherwise stated thefollowing standard protocol was used:

1) Swelling of the NovaPEG Rink Amide resin in CH₂Cl₂ for 2 hours.Deprotection of the resin is not required as it is sold withoutprotecting groups.

2) Resin is washed with a steady flow of DMF (30 seconds) followed by asteady flow of with CH₂Cl₂ (30 seconds).

3) Activation of PNA monomer (5 equivalents with respect to total activesites on the resin) occurred by addition of 4.5 equivalentsN,N,N,N-Tetramethyl-O-(7-azabenzotriazol-1-yl)uroniumhexafluorophosphate (HATU) (slightly less equivalents were used toensure total activation of monomer with HATU and to prevent occurrenceof unreacted HATU from reacting with resin amine or unprotected aminesin the growing PNA sequence. Unprotected amines can form a guanidinemoiety with HATU that blocks further elongation) and 10 equiv. ofdiisopropylethylamine (DIPEA) in DMF. The final concentration of themonomer was 0.2M in DMF. Monomer activation was allowed to proceed for10 minutes before being added to the resin.

4) A steady stream of N₂(g) was bubbled through the peptide synthesisvessel while coupling occurred. Coupling time was 60 minutes.

5) Upon completion of coupling, the activating solution was vacuumed offthe resin, and the resin was washed with a steady stream of DMF for 30seconds (3 times), followed by CH₂Cl₂ for 30 seconds (3 times). Nocapping steps were necessary for the PNA sequences chosen.

6) Deprotection of the Fmoc was done using a solution of 20% piperidinein DMF for 5 minutes.

7) Steps 2-6 were repeated until chain length was complete.

8) Following the removal of the final Fmoc group, and subsequentwashings of the resin with DMF and CH₂Cl₂, the HATU-activated carboxylicacid-substituted norbornene (4) was added and a steady stream of N₂(g)bubbled through the vessel for 60 minutes (the carboxylicacid-substituted norbornene was activated using the same protocol usedfor the PNA monomers in step 3).

9) Upon completion of coupling, the activating solution was vacuumed offthe resin, and the resin was washed with a steady stream of DMF for 30seconds (3 times), followed by CH₂Cl₂ for 30 seconds (3 times).

10) Step 8 was repeated (carboxylic acid-substituted norbornene (1)coupling to the resin).

11) Upon completion of coupling, the activating solution was vacuumedoff the resin, and the resin was washed with a steady stream of DMF for30 seconds (3 times), followed by CH₂Cl₂ for 30 seconds (3 times).

12) The resin was dried under vacuum for several hours.

13) The removal the Bhoc protecting groups and cleavage from the resinwas accomplished by treatment with a solution of TFA:m-cresol (80:20)for 90 minutes. The cleavage was performed in a polypropylene Poly-PrepChromatography Column. After cleavage, the TFA:cresol solution wasseparated from the resin by centrifugation. The TFA:cresol solution wasthen evaporated until near dryness by applying a stream of air to thesolution for several hours.

14) The crude PNA-norbornene oligomer crashed out of the TFA:cresolsolution upon addition of 5 equivalents of diethyl ether with respect tothe TFA:m-cresol solution, yielding an off-white powdery solid.

15) Reverse-phase preparatory HPLC was used to purify all sequences, andmasses were confirmed by Matrix Assisted Laser Desorption-Time of Flight(MALDI-TOF)

3. HPLC Purification of PNA Sequences

RP-HPLC analysis of PNA was performed using 0.1% TFA/H₂O as solvent A,and 0.1% TFA/CH₃CN as solvent B. Gradient: 0% solvent B in 2 min, 0% to5% solvent B in 3 min, 5% to 20% solvent B in 10 min, and 20% to 100%solvent B in 20 min.

4. Homopolymer Synthesis Via Ring-Opening Metathesis Polymerization(ROMP)

PNA-Nb monomer (CGAGTCATTT-Nb) was polymerized via ROMP using Grubbs'modified 2^(nd) generation catalyst (IMesH₂)(C₅H₅N)₂(Cl)₂Ru═CHPh in aglove box. The PNA-Nb monomer (3 mg, 1 μmol) in a J-Young NMR tube wasdissolved in 250 μL of anhydrous and degassed DMF-d₇. The tube wasremoved from the glove box and a H NMR spectrum (t=0) was taken. Thetube was returned to the glove box and the catalyst (0.2 μmol or 0.1μmol) was added to the reaction solution. ¹H NMR spectra were recordedat the indicated time points until consumption of the olefin. It wasobserved that as the PNA monomer olefin disappeared, the correspondingDMF-d₇ solutions became cloudy, and that the polyolefin peaks typicallyseen at ˜5.5 ppm were absent, indicating that the resultant polymer hadlimited to no solubility in DMF. In addition, the homopolymers wereinsoluble in H₂O, MeOH, and DCM solutions and had limited solubility ina solution of 0.05 M LiBr in DMF. Upon consumption of the olefin, thetube was returned to the glove box and termination agent ethyl vinylether (100 μL, excess) was added to the reaction mixture, and themixture was allowed to sit at room temperature for 20 minutes. The crudepolymer was precipitated from cold methanol and analyzed by SEC-MALS.

5. ROMP Timescale ¹H NMR for homopolyPNAs:

FIG. 5. Integrations based on 10 eq of PNA olefin at t=0. DMF residualproton is then integrated accordingly for the ensuing time points.

FIG. 6. Integrations based on 5 eq of PNA olefin at t=0. DMF residualproton is then integrated accordingly for the ensuing time points.

FIG. 7. SEC-MALS for I. The M_(n) was determined to be 13,790 with a PDIof 1.388, giving a degree of polymerization of 5 by LS, as opposed to 10by ¹H NMR. This discrepancy can be attributed to error in the assignmentof the dn/dc. The dn/dc used to calculate the M_(n) was 0.179, the knowndn/dc for polystyrene in DMF. In addition, a large LS peak can be seenat 16 minutes. This peak corresponded to a M_(n) of 7.3×10⁶ andindicated polymer aggregation in DMF. SEC-MALS for III could not beobtained due to polymer insolubility in DMF.

6. Block Copolymer Synthesis Via Ring-Opening Metathesis Polymerization(ROMP)

FIG. 8. General scheme of ROMP synthesis of block copolyPNA usingGrubb's 2^(nd) generation modified catalyst. A small aliquot of block 1was terminated using III (ethyl vinyl ether) for SEC-MALS analysisbefore adding PNA. This provided an accurate M_(n) and degree ofpolymerization for the first block. After polymerization of the PNAblock, the complete block copolymer was again analyzed using SEC-MALS.

7. Timescale ¹H NMR of ROMP of PNA Block Copolymers

FIG. 9. ¹H NMR timescale for 11. To a live catalyst on the end of apolyphenyl was added PNA (5 eq w.r.t. catalyst). The timescale shown isafter 17 hours of reaction, at which point the polymer was terminated.The integrals shown are based on the amount of addedphenyl-functionalized norbornene (35 eq. w.r.t catalyst, 5protons/phenyl plus 5 protons of phenyl alkylidene for a total of 180protons) and the amount of added PNA-Nb (5 eq w.r.t. catalyst).

Synthetic Procedure Details for ROMP of II

N-phenyl-cis-5-norbornene-exo-dicarboximide¹ (1) in a J-Young NMR tube(5 mg, 0.02 mmol) was dissolved in 250 μL anhydrous and degassed DMF-d₇in a glove box. Catalyst (IMesH₂)(C₅H₅N)₂(Cl)₂Ru═CHPh (0.411 mg, 0.56μmol) was added and the tube was removed from the glove box and ¹H NMRspectra were recorded until compete consumption of olefin. After olefinconsumption, the tube was returned to the glove box and PNA-Nb (8.2 mg,2.8 μmol, 5 equivalents w.r.t. catalyst) was added in 100 μL anhydrousand degassed DMF-d₇. The tube was removed from the glove box and ¹H NMRspectra were recorded at the indicated time points. Upon consumption ofthe olefin, the tube was returned to the glove box and termination agentethyl vinyl ether (100 μL, excess) was added to the reaction mixture,and the mixture was allowed to sit at room temperature for 20 minutes.The crude polymer was precipitated from cold methanol and analyzed bySEC-MALS.

FIG. 10. ¹H NMR timescale for ROMP of 3 identical block copolymersIV-VI. One lot of polyphenyl was distributed evenly for the synthesis ofthe 3 block copolymers. The amount of PNA-Nb added was identical (7.5 eqw.r.t. catalyst). The timepoint shown is after 12 hours of reaction, atwhich point all three polymers were terminated. The integrals shown arebased on the SLS value determined for the polyphenyl block (M_(n) was7,546 g/mol, giving a degree of polymerization of 30) and the amount ofadded PNA-Nb. Based on this value, the total degree of polymerizationshould be 37.5.

Synthetic Procedure Details for ROMP of IV-VI

N-phenyl-cis-5-norbornene-exo-dicarboximide¹ (1) was polymerized bydissolving 13.57 mg (0.05 mmol) in 200 μL DMF-d₇ and mixing with 1.3 mg(0.0017 mmol) of catalyst dissolved in 50 μL DMF. After completepolymerization (10 min), 6.4 μl of this reaction (0.044 μmol w.r.t.catalyst) was taken on and added to 1 mg (0.34 μmol) of PNA norbornylmonomer (this reaction described below). The remaining phenylhomopolymerization reaction mixture was quenched with excess ethyl vinylether, precipitated and analyzed by SEC-MALS (DP=30).

For incorporation of PNA norbonyl monomer as the second block in aphenyl-PNA block copolymer, 3.5 mg (1.2 μmol) PNA monomer was dissolvedwith heating in 15 μL DMSO-d₆ and diluted to 65 μL with 50 μL of DMF-d₇to yield a stock solution of PNA monomer at a concentration of 18.5 mM.18.6 μL of this PNA stock solution (1 mg, 0.34 μmol) was added to 6.4 μlof phenyl homopolymer solution with live ruthenium catalyst (asdescribed above) for a total reaction volume of 25 μL. This exactprotocol was followed for 3 identical 25 μL reactions. Each of the threereactions was heated in a glass HPLC insert at 40° C. on a heat blockwith sand used to facilitate efficient heat transfer between the heatingblock and the glass HPLC vial insert. After 1 hour of heating, all threereactions were removed from heat and subsequently diluted to 80 μL totalvolume with DMF-d₇ and added to a 3 mm O.D. NMR tube via a heat-pulledglass pipette in order to provide enough volume for NMR analysis whilekeeping the concentration at a maximum. After 12 hours, an NMR spectrumwas acquired for each of the three samples. Following this, the tube wasreturned to the glove box and termination agent ethyl vinyl ether (100μL, excess) was added to the reaction mixture, and the mixture wasallowed to sit at room temperature for 20 minutes. The crude polymer wasprecipitated from cold methanol and analyzed by SEC-MALS.

FIGS. 11A-D. ¹H NMR timescale for VI I-X.

The timescale shown is after 12 hours of reaction, at which point allpolymers were terminated. The integrals shown are based on the SLS valuedetermined for the first block (M_(n) was 11,900 g/mol, giving a degreeof polymerization of 33 for peg, M_(n) was 13,430 g/mol, giving a degreeof polymerization of 41 for NR4, and the M_(n) was 8,827 g/mol, giving adegree of polymerization of 35 for ph) and the amount of added PNA-Nb.Based on this value, the total degree of polymerization should be 45 foreach block copolymer.

ROMP Conditions for VII-X

N-phenyl-cis-5-norbornene-exo-dicarboximide (1) (3.47 mg, 13.7 mole),2-(2,5,8,11-tetraoxatridecan-13-yl)-3a,4,7,7a-tetrahydro-1H-4,7-methanoisoindole-1,3(2H)-dione(2) (4.8 mg, 13.7 μmol), andN-benzyl-2-(1,3-dioxo-1,3,3a,4,7,7a-hexahydro-2H-4,7-epoxyisoindol-2-yl)-N,N-dimethylethan-1-aminium(3) (4.47 mg, 13.7 μmol) in 85.5 μL DMF-d₇ were polymerized by mixingwith 0.274 mg (0.38 μmol) of catalyst (14.5 μL of a 0.026 M soln. inDMF-d₇) for a total volume of 100 μL DMF-d₇. After completepolymerization, 10 μl of this reaction (0.038 μmol w.r.t. catalyst) wastaken out and added to 1 mg (0.34 μmol) of PNA norbornyl monomer (thisreaction described below). The remaining polymerized first blockreaction mixture was quenched with excess ethyl vinyl ether,precipitated and analyzed by SEC-MALS.

For incorporation of PNA norbonyl monomer as the second block in aphenyl-PNA block copolymer, 4.2 mg (1.4 μmol) PNA monomer was dissolvedwith heating in 25 μL DMSO-d₆ to yield a stock solution of PNA monomerat a concentration of 57.5 mM. 5.95 μL of this PNA stock solution (1 mg,0.34 μmol) was added to 10 μl of phenyl homopolymer solution with liveruthenium catalyst (as described above) for a total reaction volume of15.95 μL. After 12 hours at r.t., all four reactions were diluted to 90uL total volume with DMF-d₇ and added to a 3 mm O.D. NMR tube via aheat-pulled glass pipette in order to provide enough volume for NMRanalysis while keeping the concentration at a maximum.

FIGS. 12A-12B. FIG. 12A) SEC-MALS of II. SEC-MALS of the polyphenylblock was not observed due to an insufficient aliquot removal of thepolyphenyl block. However, complete consumption of the phenyl-norbornenebackbone was observed by ¹H NMR. After complete polymerization of theblock copolymer, the M_(n) of the total polymer (28,270) and the PDI(1.035) were determined by SEC-MALS. M_(n) gives a degree ofpolymerization of 6 for PNA (mass fraction of this RI peak is 97.9%).The phenyl-norbornene monomer (1) was added in 35 equivalents w.r.t. toPNA. A large LS peak, but small RI (corresponding to a mass fraction of2.1%) peak can be seen at 20 minutes. This peak corresponded to a M_(n)of 3.3×10⁵ and indicated block copolymer aggregation in DMF. FIG. 12B)¹HNMR for the polymerization of the phenyl-norbornene block of ph₃₅PNA₅.The red star indicates the olefin peak at 6.32 ppm that is observed todisappear upon polymerization. The peak corresponds to the 2 protonsalso indicated by a star in the chemical structure.

FIGS. 13A-13D. SEC-MALS of polyphenyl block of as well as SEC-MALS of 3identical block copolymers of PNA (IV-VI). After complete polymerizationof phenyl-norbornene monomer (1), a predetermined volume was removedfrom the solution and terminated with ethyl vinyl ether to obtain theM_(n) of the polyphenyl block, which was determined to be 7,546, givinga degree of polymerization of 30 for the phenyl block, with a PDI of1.03. After termination of the complete block copolymer, the M_(n) wasdetermined for all 3 identical polymers and for all peaks that could beanalyzed. FIG. 13A) M_(n) was 7.7×10⁶, PDI 2.06, and mass fraction was26% for peak between 15.4-20.5 min by RI. M_(n) was 14,810, PDI 1.54,and mass fraction was 48% for peak between 20.6-24.8 min by RI. Thispeak was used to determine degree of polymerization for the PNA, as itmost likely corresponds to the disaggregated block copolymer. M_(n) was1,860, PDI 1.41, and mass fraction was 26% for peak between 25-26.5 minby RI, corresponding to unpolymerized PNA-Nb (2922 g/mol). FIG. 13B)M_(n) was 1.5×10⁶, PDI 1.45, and mass fraction was 10% for peak between16-20.6 min by RI. M_(n) was 18,700, PDI 1.34, and mass fraction was 54%for peak between 20.9-24.9 min by RI. This peak was used to determinedegree of polymerization for the PNA, as it most likely corresponds tothe disaggregated block copolymer. M_(n) was 3,850, PDI 1.11, and massfraction was 36% for peak between 25.1-27.3 min by RI, corresponding tounpolymerized PNA-Nb (2922 g/mol). FIG. 13C) M_(n) was 1.6×10⁵, PDI2.36, and mass fraction was 16% for peak between 16.9-19.5 min by RI.M_(n) was 11,210, PDI 1.25, and mass fraction was 52% for peak between19.9-24.2 min by RI. This peak was used to determine degree ofpolymerization for the PNA, as it most likely corresponds to thedisaggregated block copolymer. M_(n) was 1,661, PDI 1.23, and massfraction was 32% for peak between 25-26.5 min by RI, corresponding tounpolymerized PNA-Nb (2922 g/mol). FIG. 13D) Expanded view of LS peaksfor all 3 block copolymers.

FIGS. 14A-14B SEC-MALS of polyphenyl block as well as SEC-MALS of VII.FIG. 14A) After complete polymerization of phenyl norbornene monomer(1), a predetermined volume was removed from the solution and terminatedwith ethyl vinyl ether to obtain the M_(n), which was determined to be8,827, giving a degree of polymerization of 35, with a PDI of 1.02.After termination of the complete block copolymer, the M_(n) wasdetermined for all peaks that could be analyzed. M_(n) was 6.2×10⁵, PDI2.66, and mass fraction was 25% for peak between 14.6-20.2 by RI. M_(n)was 16,840, PDI 1.33, and mass fraction was 51% for peak between20.7-25.1 min by RI. This peak was used to determine degree ofpolymerization for the PNA, as it most likely corresponds to thedisaggregated block copolymer. The M_(n) for peak between 25.6-27.3 minby RI could not be determined due to inadequate LS. The mass fractionfor this peak was 24%. FIG. 14B) Expanded view of RI peak showing 3populations.

FIGS. 15A-15B SEC-MALS of polyphenyl block as well as SEC-MALS of VIII.FIG. 15A) After complete polymerization of phenyl norbornene monomer(1), a predetermined volume was removed from the solution and terminatedwith ethyl vinyl ether to obtain the M_(n), which was determined to be8,827, giving a degree of polymerization of 35, with a PDI of 1.02.After termination of the complete block copolymer, the M_(n) wasdetermined for all peaks that could be analyzed. M_(n) could not bedetermined for the LS peak between 13.9-19.3 min due to a lack of RI.M_(n) for the peak between 22-25 min by RI could not be determined dueto inadequate LS. This peak had a mass fraction of 45%. The M_(n) forpeak between 25.2-27.3 min by RI could not be determined due toinadequate LS. The mass fraction for this peak was 55%. FIG. 15B)Expanded view of RI peak showing 2 populations.

FIGS. 16A-16B SEC-MALS of first block as well as SEC-MALS of IX. FIG.16A) After complete polymerization of peg-norbornene monomer (2), apredetermined volume was removed from the solution and terminated withethyl vinyl ether to obtain the M_(n) of the polypeg block, which wasdetermined to be 11,900, giving a degree of polymerization of 33 for thepeg block, with a PDI of 1.03. After termination of the complete blockcopolymer, the M_(n) was determined for all peaks that could beanalyzed. M_(n) was 2.5×10⁶, PDI 1.29, and mass fraction was 12% forpeak between 15-20 by RI. Despite showing multiple peaks by RI, theM_(n) could not be determined for any other peaks due to lack of an LSpeak. The mass fraction for RI peak between 20.5-25 min was 61%, and 27%for the peak between 25.15-27.7 min. FIG. 16B) Expanded view of RI peaksshowing 3 populations.

FIGS. 17A-17B SEC-MALS of quaternary amine block as well as SEC-MALS ofX. FIG. 17A) After complete polymerization of quaternaryamine-norbornene monomer (3), a predetermined volume was removed fromthe solution and terminated with ethyl vinyl ether to obtain the M_(n),which was determined to be 13,430, giving a degree of polymerization of41, with a PDI of 1.1. After termination of the complete blockcopolymer, the M_(n) was determined for all peaks that could beanalyzed. M_(n) was 4.6×106, PDI 1.05, and mass fraction was 18% forpeak between 13.9-19.8 by RI. Despite showing multiple peaks by RI, theM_(n) could not be determined for any other peaks due to lack of an LSpeak. The mass fraction for RI peak between 20.6-24.8 min was 43%, and39% for the peak between 25-27.1 min. FIG. 17B) Expanded view of RIpeaks showing 3 populations.

8. PNA-Polymer Micelle Formation

Synthesis

6 mg of 11 was dissolved into DMSO at a concentration of 2 mg/ml. Thissolution was transferred to 3,500 MWCO snakeskin dialysis tubing (ThermoScientific) and 3 ml H₂O was added. The resulting solution was dialyzedagainst 1.0 L of Nanopure H₂O for 3 days, with the H₂O being changeddaily. UV confirmed the concentration of the final solution and aspeedvac was used to concentrate solutions to ˜0.1 mg/ml. Nucleic acidconcentrations were determined by UV absorbance at 260 nm using a ThermoScientific NanoDrop 2000c spectrophotometer. An extinction coefficientof 99,200 L/mol⁻¹cm⁻¹ was used. This coefficient was calculated as theextinction coefficient of the entire sequence.

9. TEM

Copper grids (formvar/carbon-coated, 400 mesh copper, Ted Pella #01754)were prepared by glow discharging the surface at 20 mA for 1.5 minutesfollowed by treatment with 3.5 μL 250 mM MgCl₂ in order to prepare thesurface for PNA nanoparticle adhesion. The MgCl₂ solution was wickedaway with filter paper and 3.5 μL of PNA nanoparticle (ca 100 μM PNA)solution was deposited on the grid surface. This solution was allowed tosit for 5 minutes before being washed away with 4 drops of glassdistilled H₂O and subsequent staining with 3 drops of 1% w/w uranylacetate. The stain was allowed to sit for 30 seconds before wicking awaywith filter paper. All grid treatments and sample depositions were onthe dark/shiny/glossy formvar-coated face of the grid (this side face upduring glow discharge). Samples were then imaged via TEM.

FIGS. 18A-18D. FIG. 18A) After dialysis into H₂O from DMSO, II formsnanoparticles that by DLS are aggregates on the size order of 50 nm indiameter. FIG. 18B) Autocorrelation function for nanoparticles formedfrom copolyPNA-3. FIG. 18C) Negative-stain TEM showing nanoparticles onthe order of 10-30 nm. The majority of the material, once dried, showedno particle aggregation. FIG. 18D) Negative stain TEM showing azoomed-out section of the grid.

10. DNA Melting Temperature Analysis

Melting temperature analysis were performed by heating each sample from20° C. (20 minute equilibration time) to 90° C. using a temperaturegradient of 1° C./minute. Melting temperatures were calculated as firstderivatives of the curve. Nanoparticles formed from II, renamed asPNA-NP, were at a concentration of 1 μM in water. The mixture was madeby adding Dulbecclo's 1×PBS to the nanoparticles followed by 100 nM-1 μMof the complementary DNA in H₂O. Final concentration of NaH₂PO₄ is 6.7mM, NaCl is 113 mM, KCl is 22.2 mM, and KH₂PO₄ is 1.46 mM, all in atotal volume of 50 μL. Annealing was done at room temperature for 2hours. The sample was refrigerated at 8° C. for 15 minutes, afterannealing, and subsequently analyzed.

FIGS. 19A-19C. FIG. 19A) Raw T_(m) data for PNA-PN and complementary DNAsequence. Several buffers were tested to determine ideal conditions forhybridization between PNA-NP and its DNA complement at room temperature.1 μM PNA-NP with 100 nM complementary DNA in PBS was subsequentlychosen. FIG. 19B) A 10-pt FFT filter was applied to the raw data. FIG.19C) Derivation plots for each T_(m) curves showing 57.8° C. average forPNA-NP and complementary DNA.

FIGS. 20A-20C. FIG. 20A) Raw T_(m) data for PNA-NP and complementaryDNA, as well as non-complementary DNA sequence. The T_(m) for thecorresponding PNA sequence (identical to PNA that was polymerized) andcomplementary DNA was also determined. FIG. 20B) A 10-pt FFT filter wasapplied to the raw data. FIG. 20C) Derivative plots for each of theT_(m) curves showing a 7.3° C. increase for PNA-NP over its identicalPNA sequence. While a derivative can be taken of the T_(m) curvecorresponding to PNA-NP and its non-complementary sequence, the curveitself is more indicative of non-specific binding, as is implied by thebroad derivative, and the almost linear T_(m).

FIGS. 21A-21B. FIG. 21A) Raw T_(m) data for PNA-NP without complementaryDNA and complementary DNA without PNA in PBS showing no melting. FIG.21B) A 10-pt FFT filter was applied to the raw data.

11. Molecular Dynamics (MD) Simulations

A single PNA-polynorbornyl unimer was initially built in fully extendedconformation with GaussView software [GaussView, Version 5, Dennington,R.; Keith, T.; Millam, J. Semichem Inc., Shawnee Mission Kas., 2009]. 60identical unimers, first relaxed in vacuum for 0.3 ns at the temperatureof 300 K, were spherically distributed in space, with PNA ends orientedtowards the outside. The resulting PNA-NP of 60 unimers was thenimmersed in a cubic (TIP3P) water box with solvate plugin in VIVID[Humphrey, W.; Dalke, A.; Schulten, K. “VMD—Visual Molecular Dynamics”,J. Molec. Graphics, 1996, 14, 33-38]; water molecules present within a65 Å radius of PNA-NP center were deleted. The resulting unit cell withsolvated 60-monomer PNA-NP contained 2,122,554 atoms.

MD simulations of solvated PNA-NP were performed with NAMD2 software[Phillips, J. C.; Braun, R.; Wang, W.; Gumbart, J. C.; Tajkhorshid, E.;Villa, E.; Chipot, C.; Skeel, R. D.; Kale, L.; Schulten, K. Journal ofComputational Chemistry, 2005, 26, 1781-1802], where the molecules weredescribed using the CHARMM force field. The parameters for the unimerunits (PNA, norbornyl) were obtained by analogy to molecules alreadyparametrized in the CHARMM forcefield, using the ParamChem Server[Vanommeslaeghe, K.; Hatcher, E.; Acharya, C.; Kundu, S.; Zhong, S.;Shim, J.; Darian, E.; Guvench, O.; Lopes, P.; Vorobyov, I.; MacKerellJr., A. D. J. Comput. Chem. 2010, 31, 671-690; Vanommeslaeghe, K.;MacKerell Jr., A. D. J. Chem. Inf. Model. 2012, 52, 3144-3154;Vanommeslaeghe, K.; Raman, E. P.; MacKerell Jr., A. D. J. Chem. Inf.Model. 2012, 52, 3155-3168]. All simulations were performed in NpTensemble using periodic boundary conditions, at a constant temperatureT=300 K, a Langevin constant γ_(Lang)=0.001 ps⁻¹ (to ensure fastdynamics), and at a constant pressure p=1.01325 bar. The particle-meshEwald (PME) method [Darden, T.; York, D.; Pedersen, L. J Chem. Phys.1993, 98, 10089] was used for evaluation of long-range Coulombicinteractions. The timestep was set to 1.0 fs, and long rangeinteractions were evaluated every 1 (van der Waals) and 2 timesteps(Coulombic).

In the prepared system, water was minimized for 10 ps around the fixedPNA-NP, then for additional 8 ps around the constrained PNA-NP. Thewhole system was then heated to the temperature of 300 K andequilibrated at this temperature for 16 ns without constraints.

Partial Atomic Charges

CHARMM parameters for all atoms were prepared by analogy to knownmolecules, using the ParamChem Server [4-6]. The obtained partialcharges were slightly modified to ensure that the unimer had no netcharge. Below are the partial atomic charges for each PNA base, a unitof the PNA peptide chain, and a unit of the hydrophobic chain.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, one of skill in the art will appreciate that certainchanges and modifications may be practiced within the scope of theappended claims. In addition, each reference provided herein isincorporated by reference in its entirety to the same extent as if eachreference was individually incorporated by reference.

What is claimed is:
 1. A graft polymer comprising a linear backbonecovalently bound to a plurality of oligonucleotide branches; wherein thegraft polymer has the formula:R¹—[M(O)]_(n)—R² wherein, n is an integer from 2 to 1000; M is apolymerized ROMP polymerizable monomer; O is an oligonucleotidecovalently attached directly to M or to M through a covalent linker; andR¹ and R² are terminal polymer moieties, wherein 100% of the polymerizedROMP polymerizable monomers are each individually attached directly tothe oligonucleotide or to the oligonucleotide through a covalent linker,and wherein the polymerizable monomer isN-substituted-5-norbornene-2,3-dicarboximide, wherein the substitutioncomprises the oligonucleotide.
 2. The graft polymer of claim 1, whereinsaid oligonucleotide comprises at least 3 nucleobases and at least 2different nucleobases.
 3. The graft polymer of claim 1 comprising atleast 3 oligonucleotide branches.
 4. The graft polymer of claim 1,wherein R¹ and R² each independently comprise a substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl or substituted orunsubstituted heteroaryl.
 5. The graft polymer of claim 1, wherein M(O)is wherein,

L¹ is independently a bond, —O—, —NH—, —COO—, —S—, —SO₂—, —SO₃—, —SO₄—,—SO₂NH—, —NHC(O)—, —C(O)NH—, —NHC(O)O—, substituted or unsubstitutedalkylene, substituted or unsubstituted heteroalkylene, substituted orunsubstituted cycloalkylene, substituted or unsubstitutedheterocycloalkylene, substituted or unsubstituted arylene, orsubstituted or unsubstituted heteroarylene; and R⁴ is theoligonucleotide.
 6. The graft polymer of claim 5, wherein R¹ and R² eachindependently comprise a substituted or unsubstituted alkyl, substitutedor unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl or substituted or unsubstituted heteroaryl.
 7. Thegraft polymer of claim 6, wherein the oligonucleotide comprises at least3 nucleobases and at least 2 different nucleobases.